This standard specifies the electrophoresis method and judgment criteria for the detection of biochemical markers of inbred mice and rats using cellulose acetate membrane (plate). This standard is applicable to any strain of inbred rats and mice. GB/T 14927.1-2001 Laboratory Animals - Detection Methods of Biochemical Markers of Inbred Mice and Rats GB/T14927.1-2001 Standard Download Decompression Password: www.bzxz.net

ICS 65. 020.30
National Standard of the People's Republic of China
GB/T 14927. 1—2001
Laboratory animal
Genetic monitoring : methods for biochemicalmarkers of inbred mice and rats2001-08-29 Issued
General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China
Implementation on 2002-05-01
GB/T14927.1—2001
This standard is a revision of GB/T14927.1-1994 "Laboratory animal-Genetic monitoring : methods for biochemicalmarkers of inbred mice and rats". This standard promotes the electrophoresis results of the biochemical marker detection method of inbred mice and human mice to be as vivid as possible, so as to facilitate the application of experimental results comparison. The biochemical sites in the biochemical marker detection method of inbred mice are adjusted, the biochemical site of major urinary protein-1 (Mup1) with poor operability is cancelled, and the alkaline phosphatase-1 (Akp1) site with strong operability and easy promotion and application is added. The esterase-1 (Es1) biochemical site commonly used in the national standard is added.
This standard has revised the reagent name, writing specifications, etc. This standard and its supporting standards will replace CB/T14927.1-1994 from the date of implementation. This standard is proposed and managed by the Ministry of Science and Technology of the People's Republic of China. The drafting unit of this standard: China Laboratory Animal Society. The main drafters of this standard: Xing Ruichang, Liu Shuanghuan, Shen Jie, Li Shanru, Bai Qinhua, Zhang Ruizhong. This standard was first issued in January 1994.
1 Scope
National Standard of the People's Republic of China
Laboratory Animals
Genetic Monitoring: Methods for Biochemical Markers of Inhred Mice and RatsGB/T 14927. 1 —2001
Replaces GB/T14927.1--- 1994
This standard specifies the electrophoresis method and judgment criteria for the detection of biochemical markers of inbred mice and human mice on cellulose acetate membrane (plate). This standard is applicable to any strain of inbred rats and mice. 2 Definitions
This standard adopts the following definitions.
2.1 Biochemical Markers Biochemical markers referred to in this standard refer to markers that indicate genetic characteristics and are identified by biochemical methods. In mice and rats, they are mostly some isozymes and isomers.
2.2 Biochemical genetic profile: The biochemical genetic profile referred to in this standard refers to the summary of phenotypic data of multiple biochemical genetic markers of various inbred strains, which reflects the genetic characteristics of all strains to a certain extent.
2.3 Homozygosity
Homozygosity referred to in this standard refers to the state of having the same genes in the relative positions of homologous chromosomes. Inbred animals have homozygosity at most loci through continuous inbreeding, and the offspring produced by mating between any individuals in a strain are also homozygous. 2.4 Isogenicity
Isogenicity referred to in this standard refers to the fact that all individuals in an inbred strain are genetically homologous. Therefore, skin and tumor transplants between any individuals in the same strain are not regarded as alien and rejected. If the genes of inbred animals are tested, the genotypes of different individuals in a strain are completely consistent.
2-5 Individuality
Individuality in this standard means that for the entire inbred animal, each strain is genetically unique, which is manifested in a wide range of characteristics, such as biochemical genetic profiles. Most inbred strains can be distinguished from each other by their respective biochemical genetic profiles. 3 Principle of the method
There are some isozymes and isomeric proteins in rats and mice. They can be distinguished by electrophoresis based on the different charges they carry in a specific electric field, and their genotypes can be inferred based on the electrophoretic band type, that is, the phenotype of the protein, to establish the genetic profiles of various inbred strains and conduct regular quality monitoring on them. Approved by the General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China on August 29, 2001 and implemented on May 1, 2002
Equipment and materials
Normal pressure electrophoresis instrument.
Cellulose acetate membrane electrophoresis tank.
Electrophoresis spotting device.
Cellulose acetate membrane.
Cellulose acetate plate.
4℃, -20℃ refrigerator.
Low-temperature high-speed centrifuge.
Oscillator,
GB/T14927.12001
4.9 Tissue homogenizer (2 mL or 5 mL) or tissue homogenizer. Anticoagulation capillary. Anticoagulation capillary aspirator. 4.10
Anticoagulation capillary centrifuge
Ear suction bulb.
Small grinding wheel.
Dissection board, surgical scissors
Bamboo tweezers.
S water-soluble pot, 37
4.17 glass blood
4.186cm×8
ordinary filter paper
analytical balance
4.21 pH meter.
4.22 Ultraviolet monitoring #
4.23 Common laboratory chemical reagents
Boric acid
Glycine
Chinese name (or abbreviated name
aminomethane
tetraacetic acid
Citrate fcid
Mg(C,HO)
Ponceau S
Citric acid
Disodium hydrogen phosphate
Potassium dihydrogen phosphate
Sodium hydroxide
·Sodium acetate anhydrous
Agar powder
Magnesium acetate
Manganese chloride
Ponceau red-S
AR or.C, P.
AR or. CP
AR or. CP
ARor. CP
AR or. CP
AR or.CP
AR or.CP
AR or.CP
AR or.CP
AR or. CP
A, R. or. CP
CCI,COOH
English name (or write)
(HO)CH.(COOH)SO.H.2H.O
Barbital
Sodium harbital
NA,FDTA
K,[Fe(CN)
Acctonc
Glucose-1-phosphate
Cystarnine
1,4-dithiohreitol(1
β-naphthyl acetat
4-methyl-umbel
g-Naphthyl acie
D-fructose-6
Thiazalyl blue
g-uicouinami
N-methyl ph
DL-lsaitrie
Glucse-1,6
D-glucose-6-p
Acetate
hosplate
zolim bromit
ine dinucleot
GB/T 14927.12001
Chinese name
Blue chloroacetic acid
Sulfosalicylic acid
Barbital
Barbital sodium
Magnesium chloride
Ethylenediaminetetraacetic acid disodium
Ferric chloride
Potassium ferrocyanide
Hydroxy
B-acetic acid
4-methyl
β-phosphoric acid
D-button
Hydrogen salt
Baohua methyl sulfate
ummethyl sulfate(PMs)
isodium salt
Glucose-i-phos
Malic acidl
Fast blue RR sal
5 Biochemical marker detection methods
5.1. Preparation of electrophoresis samples
5. 1. 1 Plasma
dehydrogen
methoxythiophene
isocitric acid
firm blue
diphosphodiester
acid deaminase
ARor.CP
AR or. CF
ARor:CP
AR or, C, P.
ARor.CP
ARor.CP
ARor.CP
ARor.CP
ARor.CP
ARor.CP
AR ar.CP
Import or domestic
Escape from the export or domestic
Import or domestic
Import or domestic
Organize the export or domestic
Organize
Import or domestic
Import or domestic
Import and domestic
Import and domestic
Import and domestic
Import and domestic
Import and domestic
Import and domestic
Import and domestic
Escape from the export of our country
Import or domestic
Import or domestic
Import or domestic
Use anticoagulant capillary tube to perform orbital blood sampling, 500 r/min, centrifuge for 5xi to separate plasma and blood cells, and aspirate plasma for use. 5.1.2 Hemolysin
Add distilled water (1:4 standard) to the red blood cells from which the plasma has been removed for 1 minute, and a red transparent liquid will be formed, which is the hemolysin. 5.1.3 Supernatant of tissue homogenate
The animals were killed by cervical dislocation, and the laparotomy was performed. One kidney and one liver lobe were taken from mice; one kidney, 3 cm small intestine, one testis were taken from rats, and one lung was taken from the chest. An appropriate amount of pre-cooled distilled water was added to each of them. The ratio of distilled water to tissue was generally 2:1 (V/W). ). Homogenize with a vertical homogenizer. Place the homogenate in a low-temperature high-speed centrifuge and centrifuge at 16000r/rmin for 30min. Use a pipette to draw the supernatant and store it in a small test tube for later use.
5.1.4 Sample preservation
The above-prepared samples are all fresh for use. They can only be stored for one day in an ordinary box and no more than one month in a low-temperature refrigerator. 5.2 Electrophoresis steps
5.2.1 Without membrane
GB/T 14927.1--2001
Gently immerse the cellulose acetate membrane (plate) in the corresponding electrophoresis buffer. Avoid bubbles on the membrane (plate) when immersing. 5.2.2 Spotting
Take out the soaked membrane (plate), dry it with filter paper, place the cellulose membrane side up, flat on the spotting plate, take the numbered sample pre-set in the sample slot with the spotter, and spot it on the membrane (plate). The spotting volume is 0.3μI. To increase the sample volume on the membrane (plate), repeat the spotting. The optimal spotting volume should not exceed (. 9 μL (three times).
5.2.3 Electrophoresis
Use a pencil to mark the origin and the direction of electrophoresis on the membrane (plate), quickly place the membrane (plate) on the paper bridge of the electrophoresis tank that has been pre-placed with the buffer solution, cover the electrophoresis tank, and turn on the power supply. The electrophoresis conditions are as shown in Chapters 6 and 7 of this standard. 5.3 Staining
5.3.1 Protein staining method
After the electrophoresis is completed, take out the membrane (plate) and place it in 0.2% Ponceau red staining solution. After 10~15 minutes, take it out with bamboo tweezers and replace it with 7% acetic acid for decolorization until the electrophoresis zone is clearly visible.
5.3.2 Enzyme color plate method
Applicable to cellulose acetate membrane.
5. 3.2. 1 Mix the enzyme coloring solution (see Appendix B) freshly. Add 3~4 mL of 2% hot agar, mix quickly, and invert it on a 6 cm×8 5.3.2.2 After electrophoresis, remove the membrane and stick the sample surface on the enzyme color plate. Be careful to remove all bubbles between the membrane and the enzyme color plate, but do not move the position of the membrane.
5.3.2.3 Move the color plate with the membrane to a 37℃ incubator for insulation until the enzyme zone is clearly visible. 5.3.2.4 Remove the colored membrane and immerse it in 5%~7% acetic acid to terminate the reaction. 5.3.3 Agar overlay method
Applicable to cellulose acetate.
5.3.3.1 After electrophoresis, remove the cellulose acetate plate. 5.3.3.2 Quickly mix the fresh mixed enzyme color solution (see Appendix B) with 2~3mL 2% hot agar and evenly spread it on the horizontal cellulose acetate plate
5.3. 3.3 After the agar is cooled and fixed, move the cellulose acetate plate to a 37℃ incubator until the enzyme zone is clearly visible. 5.3.3.4 Place the cellulose acetate plate in 7% acetic acid to terminate the reaction. 6 Detailed Rules for Mouse Biochemical Marker Detection
6.1 Alkaline phosphatase-1 (Akpl) Chr, 16.1.1 Sample: kidney plasma, 0.6l.
6.1.2 Buffer: Iris-Citrate pH 8.3 See A9 in Appendix A (Standard Appendix). 6.1.3 Electrophoresis support: cellulose acetate membrane (plate). 6.1.4 Electrophoresis conditions: voltage = 200 V, time = 40) min, moving direction from negative electrode to positive electrode. 6.1.5 Staining method: enzyme color plate method.
6.1.6 Staining solution: See B10 in Appendix B (standard appendix). 6.1.7 Standard control;
C57BT./6J
6.1.8 Judgment method: Read the band pattern with reference to the above control animals and compare it with Appendix C (standard appendix). 6.1.9Ap1 Electrophoresis result pattern diagram, see Figure 1, 4
GB/T 14927. 1—2001
Figure 1A1 Electrophoresis result pattern diagram
6.2 Carbonic anhydrase-2 (Car2) Chr. 36.2.1 Sample: hemolysin, 0.3 μ.
Buffer NaC,H,(), EDTA pHS. 4 See AI in Appendix A (standard appendix). 6.2.2
6.2.3 Electrophoresis support: cellulose acetate membrane. 6.2.4
Electrophoresis conditions: voltage = 240V, time = 10min, moving direction from positive pole to negative pole. 6.2. 5
Staining method: protein staining method.
6.2.6 Staining solution: see B6 in Appendix 3 (Standard Appendix). 6.2.7 Standard control:
C57BL/6J
DBA/2J
6.2.8 Judgment method: refer to the above control animals to read the band pattern and compare it with Appendix C (Standard Appendix). 6.2.9 Cur2 electrophoresis result pattern diagram, first Figure 2. h
Figure 2Car2 electrophoresis result pattern
6.3 Kidney catalase-2 (Ce2) Chr.76.3.1 Sample: kidney slurry.0.3l..
6.3.2 Buffer: Tris-citrate pH7.6 See A3.6 in Appendix A (Standard Appendix) 6.3.3 Electrophoresis support: cellulose acetate membrane. 6.3.4 Electrophoresis conditions: voltage = 200V, time = 25min, moving direction from negative electrode to positive electrode. 6.3.5 Staining method: enzyme color plate method.
GB/T 14927.1--2001
6.3.6 Staining solution: See Appendix 13 (Standard Appendix) R8. 6.3.7 Standard control:
· Fast band
6.3.8 Judgment method: Read the band pattern with reference to the above control animals and compare it with Appendix C (Standard Appendix). 6.3.9 (Ce2 electrophoresis result pattern, see Figure 3. Circle
Electrophoresis result pattern
6.4 Esterase-1 (e
6.4.1Sample:
Buffer
Electrophoresis support
Electrophoresis strip
6.4.5Staining method
Staining solution:
Standard control:
-1,Es1)Chr
.3 μt.
phate buffer pH7:
Acetate cellulose membrane.
Y=140 V, time
Color plate method.
B (Standard Appendix) Judgment method: refer to
nin, transfer the coupled animal to the positive electrode.
(Standard Appendix) for comparison
s1 Power supply result pattern diagram
6-5 Es3 (esterase-3, Es3) Chr. 1l6.5.1 Sample kidney, liver to plasma, 0.3L,
GB/T 14927.12001
6.5.2 Buffer: Tris-Glycine pII8. 9 See Appendix A (Standard Appendix) A7. 6.5.3 Electrophoresis support: cellulose acetate membrane, 6.5.4 Electrophoresis conditions: voltage = 280 V, time 28 min, the direction of movement is from the negative electrode to the positive electrode. 6.5.5 Staining method Enzyme color plate method.
6.5.6 Staining solution: See B16.5.7 Standard reference:
BALB/eJ
6.5.8 Judgment method: Refer to
6.5.9 ES3 electrophoresis result grid
6.6 Esterase-10 (estera
6.6.1 Sample: kidney and liver homogenate
6-6.2 Buffer: Tris-Glye
6.6.3 Ball support: cellulose acetate
slowest band
animal reading
as Figure 5.
10) Chr
C standard
Es3 electrophoresis result pattern
A standard in the appendix
9 grams of nitrogen
high appendix for ratio conversion
6. 6. 4 Electrophoresis conditions: voltage = 280 Time = 28 tons. Moving direction: negative electrode to positive electrode
6.6.5 Staining method: enzyme color plate method
6.6.6 Staining solution: see Appendix B (standard appendix)6.6.7 Standard control:
BALB/eJ
DBA/2J
BUB/BuJ
Slowest band
6.6.8 Judgment method: refer to [the above control animal to read the band type and compare it with Appendix C (standard appendix), 6.6.9Fs10 electrophoresis result pattern diagram, see Figure 66.7 Glucose-t-phosphate dehydrogenase-1 (glucosc.6 phosphatc rlehydrogenase-1.Gpd1) Chr.46.7.1 Sample: fresh kidney or liver homogenate, 0.9l.6.7.2 Buffer: Tris-Glycine plI8.9 See A7 in Appendix A (Standard Appendix). 6.7.3 Electrophoresis support: cellulose acetate plate (membrane). 6.7.4
GB/T 14927. 1: 2001
ES10 electrophoresis result pattern
Electrophoresis conditions: voltage = 200V, time = 50 min, moving direction from negative electrode to positive electrode. Staining method: agar overlay method.
Staining solution: See B7 in Appendix 1 (Standard Appendix). Standard control:
C57BL/6]
BALB/eJ
Slowest band
6.7.8 Judgment method: Read the band pattern of the control animal mentioned above and compare it with Appendix C (Standard Appendix). 6.7.9 Gpal1 electrophoresis result pattern, see Figure? . ■
Figure 7 Gpd1 electrophoresis result pattern
6-8 Glucosephosphateisomerase-1 (Gpil) Chr.76. 8. 1 Sample: hemolysin, 0. 3 μL. .6.8.2
Buffer: Tris-Glycine pH8. 5 See A6 in Appendix A (Standard Appendix). 6.8.3 Electrophoresis support: cellulose acetate membrane. 6.8.4
Electrophoresis conditions: voltage=200 V, time=30 min, migration direction from positive electrode to negative electrode. 6.8.5
Staining method: enzyme color plate method.
Staining solution: See B3 in Appendix B (Standard Appendix). Standard control:
BALB/cJ
C57BL/6J
GB/T 14927.1—2001
6.8.8 Judgment method: Read the band pattern with reference to the above control animals and compare it with Appendix C (Standard Appendix). 6.8.9 Gpi1 electrophoresis result pattern diagram, see Figure 8. b
Gpil electrophoresis result pattern diagram
6.9 Hemoglobin β-chain (H6b) Chr, 76.9.1 Sample: Add 1/4 volume of alkylating agent to hemolysin, see Appendix B (Standard Appendix) B9, 0.3 uL. 6.9.2 Buffer: Tris-Glycine pH8.5 See A6 in Appendix A (Standard Appendix). 6.9.3 Electrophoresis support: Cellulose acetate membrane. 6.9.4 Electrophoresis conditions: Voltage = 200V, time 30min, moving direction from negative electrode to positive electrode. 6.9.5 Staining method: Protein staining method.
6.9.6 Staining solution: See B6 in Appendix B (Standard Appendix). 6.9.7 Standard control,
C57BI./6J
BALB/eJ
6.9.8 Judgment method: Read the band pattern with reference to the above control animals and compare it with Appendix C (Standard Appendix). 6.9.9H66 electrophoresis result pattern diagram, see Figure 9. Hbb
Figure 9 Hbh electrophoresis result pattern
GB/T 14927.1-2001
6. 10 Isocitrate dehydrogenase-1 and malic enzyme-1 (Idh1 and Mex1) Chr.I
Sample: The slurry was diluted with distilled water 1:4, 0.3pL, buffer: Tris-Citrate pH7.6, see Appendix A (Standard Appendix) A3. Electrophoresis conditions: voltage = 200 V, time = 35 min, moving direction from negative electrode to positive electrode. Staining method: Enzyme color plate method, Staining solution: See Appendix B (Standard Appendix) B4 Standard control: Judgment method: Idhl and M. green Result pattern Hasphog 6.11 Phosphoglucose translocation 6.11.1 Sample: kidney plasma, Buffer: 1ris-Glycine monophosphate electrophoresis See Figure! dh2 ModdhI I and Mod1 Electrophoresis Result pattern Figure 18. Appendix A0 Broad electrophoresis Support: Cellulose acetate AG.
Electrophoresis conditions: voltage = 200 V, time threshold = 40 min, moving direction from negative electrode to positive electrode.
Staining solution: See B5 in Appendix B (standard appendix).
6.11.7 Standard control:
C57BL/61
6.11.8 Judgment method: Read the band pattern of the above control animals and compare it with Appendix C (standard appendix).
6.11.9 Pgml electrophoresis result pattern diagram, see Figure 1
6.12 Transferrin (ferransferin, 1r) Chr.96 12.1 Sample: serum, 0.3 μL
6.12.2 Buffer: Tris-Glycine pI18.5 See A6 in Appendix A (standard appendix).
6.12.3 Electrophoresis support: cellulose acetate membrane. 10
phate buffer pH7:
cellulose acetate membrane.
jade=140 V, when
color plate method.
B (Standard Appendix) Judgment method: refer to
nin, transfer the coupled animal to the positive electrode.
(Standard Appendix) for comparison
s1 Power supply result pattern diagram
6-5 Es3 (esterase-3, Es3) Chr. 1l6.5.1 Sample kidney, liver to plasma, 0.3L,
GB/T 14927.12001
6.5.2 Buffer: Tris-Glycine pII8. 9 See Appendix A (Standard Appendix) A7. 6.5.3 Electrophoresis support: cellulose acetate membrane, 6.5.4 Electrophoresis conditions: voltage = 280 V, time 28 min, the direction of movement is from the negative electrode to the positive electrode. 6.5.5 Staining method Enzyme color plate method.
6.5.6 Staining solution: See B16.5.7 Standard reference:
BALB/eJ
6.5.8 Judgment method: Refer to
6.5.9 ES3 electrophoresis result grid
6.6 Esterase-10 (estera
6.6.1 Sample: kidney and liver homogenate
6-6.2 Buffer: Tris-Glye
6.6.3 Ball support: cellulose acetate
slowest band
animal reading
as Figure 5.
10) Chr
C standard
Es3 electrophoresis result pattern
A standard in the appendix
9 grams of nitrogen
high appendix for ratio conversion
6. 6. 4 Electrophoresis conditions: voltage = 280 Time = 28 tons. Moving direction: negative electrode to positive electrode
6.6.5 Staining method: enzyme color plate method
6.6.6 Staining solution: see Appendix B (standard appendix)6.6.7 Standard control:
BALB/eJ
DBA/2J
BUB/BuJ
Slowest band
6.6.8 Judgment method: refer to [the above control animal to read the band type and compare it with Appendix C (standard appendix), 6.6.9Fs10 electrophoresis result pattern diagram, see Figure 66.7 Glucose-t-phosphate dehydrogenase-1 (glucosc.6 phosphatc rlehydrogenase-1.Gpd1) Chr.46.7.1 Sample: fresh kidney or liver homogenate, 0.9l.6.7.2 Buffer: Tris-Glycine plI8.9 See A7 in Appendix A (Standard Appendix). 6.7.3 Electrophoresis support: cellulose acetate plate (membrane). 6.7.4
GB/T 14927. 1: 2001
ES10 electrophoresis result pattern
Electrophoresis conditions: voltage = 200V, time = 50 min, moving direction from negative electrode to positive electrode. Staining method: agar overlay method.
Staining solution: See B7 in Appendix 1 (Standard Appendix). Standard control:
C57BL/6]
BALB/eJ
Slowest band
6.7.8 Judgment method: Read the band pattern of the control animal mentioned above and compare it with Appendix C (Standard Appendix). 6.7.9 Gpal1 electrophoresis result pattern, see Figure? . ■
Figure 7 Gpd1 electrophoresis result pattern
6-8 Glucosephosphateisomerase-1 (Gpil) Chr.76. 8. 1 Sample: hemolysin, 0. 3 μL. .6.8.2
Buffer: Tris-Glycine pH8. 5 See A6 in Appendix A (Standard Appendix). 6.8.3 Electrophoresis support: cellulose acetate membrane. 6.8.4
Electrophoresis conditions: voltage=200 V, time=30 min, migration direction from positive electrode to negative electrode. 6.8.5
Staining method: enzyme color plate method.
Staining solution: See B3 in Appendix B (Standard Appendix). Standard control:
BALB/cJ
C57BL/6J
GB/T 14927.1—2001
6.8.8 Judgment method: Read the band pattern with reference to the above control animals and compare it with Appendix C (Standard Appendix). 6.8.9 Gpi1 electrophoresis result pattern diagram, see Figure 8. b
Gpil electrophoresis result pattern diagram
6.9 Hemoglobin β-chain (H6b) Chr, 76.9.1 Sample: Add 1/4 volume of alkylating agent to hemolysin, see Appendix B (Standard Appendix) B9, 0.3 uL. 6.9.2 Buffer: Tris-Glycine pH8.5 See A6 in Appendix A (Standard Appendix). 6.9.3 Electrophoresis support: Cellulose acetate membrane. 6.9.4 Electrophoresis conditions: Voltage = 200V, time 30min, moving direction from negative electrode to positive electrode. 6.9.5 Staining method: Protein staining method.
6.9.6 Staining solution: See B6 in Appendix B (Standard Appendix). 6.9.7 Standard control,
C57BI./6J
BALB/eJ
6.9.8 Judgment method: Read the band pattern with reference to the above control animals and compare it with Appendix C (Standard Appendix). 6.9.9H66 electrophoresis result pattern diagram, see Figure 9. Hbb
Figure 9 Hbh electrophoresis result pattern
GB/T 14927.1-2001
6. 10 Isocitrate dehydrogenase-1 and malic enzyme-1 (Idh1 and Mex1) Chr.I
Sample: The slurry was diluted with distilled water 1:4, 0.3pL, buffer: Tris-Citrate pH7.6, see Appendix A (Standard Appendix) A3. Electrophoresis conditions: voltage = 200 V, time = 35 min, moving direction from negative electrode to positive electrode. Staining method: Enzyme color plate method, Staining solution: See Appendix B (Standard Appendix) B4 Standard control: Judgment method: Idhl and M. green Result pattern Hasphog 6.11 Phosphoglucose translocation 6.11.1 Sample: kidney plasma, Buffer: 1ris-Glycine monophosphate electrophoresis See Figure! dh2 ModdhI I and Mod1 Electrophoresis Result pattern Figure 18. Appendix A0 Broad electrophoresis Support: Cellulose acetate AG.
Electrophoresis conditions: voltage = 200 V, time threshold = 40 min, moving direction from negative electrode to positive electrode.
Staining solution: See B5 in Appendix B (standard appendix).
6.11.7 Standard control:
C57BL/61
6.11.8 Judgment method: Read the band pattern of the above control animals and compare it with Appendix C (standard appendix).
6.11.9 Pgml electrophoresis result pattern diagram, see Figure 1
6.12 Transferrin (ferransferin, 1r) Chr.96 12.1 Sample: serum, 0.3 μL
6.12.2 Buffer: Tris-Glycine pI18.5 See A6 in Appendix A (standard appendix).
6.12.3 Electrophoresis support: cellulose acetate membrane. 10
phate buffer pH7:
cellulose acetate membrane.
jade=140 V, when
color plate method.
B (Standard Appendix) Judgment method: refer to
nin, transfer the coupled animal to the positive electrode.
(Standard Appendix) for comparison
s1 Power supply result pattern diagram
6-5 Es3 (esterase-3, Es3) Chr. 1l6.5.1 Sample kidney, liver to plasma, 0.3L, bZxz.net
GB/T 14927.12001
6.5.2 Buffer: Tris-Glycine pII8. 9 See Appendix A (Standard Appendix) A7. 6.5.3 Electrophoresis support: cellulose acetate membrane, 6.5.4 Electrophoresis conditions: voltage = 280 V, time 28 min, the direction of movement is from the negative electrode to the positive electrode. 6.5.5 Staining method Enzyme color plate method.
6.5.6 Staining solution: See B16.5.7 Standard reference:
BALB/eJ
6.5.8 Judgment method: Refer to
6.5.9 ES3 electrophoresis result grid
6.6 Esterase-10 (estera
6.6.1 Sample: kidney and liver homogenate
6-6.2 Buffer: Tris-Glye
6.6.3 Ball support: cellulose acetate
slowest band
animal reading
as Figure 5.
10) Chr
C standard
Es3 electrophoresis result pattern
A standard in the appendix
9 grams of nitrogen
high appendix for ratio conversion
6. 6. 4 Electrophoresis conditions: voltage = 280 Time = 28 tons. Moving direction: negative electrode to positive electrode
6.6.5 Staining method: enzyme color plate method
6.6.6 Staining solution: see Appendix B (standard appendix)6.6.7 Standard control:
BALB/eJ
DBA/2J
BUB/BuJ
Slowest band
6.6.8 Judgment method: refer to [the above control animal to read the band type and compare it with Appendix C (standard appendix), 6.6.9Fs10 electrophoresis result pattern diagram, see Figure 66.7 Glucose-t-phosphate dehydrogenase-1 (glucosc.6 phosphatc rlehydrogenase-1.Gpd1) Chr.46.7.1 Sample: fresh kidney or liver homogenate, 0.9l.6.7.2 Buffer: Tris-Glycine plI8.9 See A7 in Appendix A (Standard Appendix). 6.7.3 Electrophoresis support: cellulose acetate plate (membrane). 6.7.4
GB/T 14927. 1: 2001
ES10 electrophoresis result pattern
Electrophoresis conditions: voltage = 200V, time = 50 min, moving direction from negative electrode to positive electrode. Staining method: agar overlay method.
Staining solution: See B7 in Appendix 1 (Standard Appendix). Standard control:
C57BL/6]
BALB/eJ
Slowest band
6.7.8 Judgment method: Read the band pattern of the control animal mentioned above and compare it with Appendix C (Standard Appendix). 6.7.9 Gpal1 electrophoresis result pattern, see Figure? . ■
Figure 7 Gpd1 electrophoresis result pattern
6-8 Glucosephosphateisomerase-1 (Gpil) Chr.76. 8. 1 Sample: hemolysin, 0. 3 μL. .6.8.2
Buffer: Tris-Glycine pH8. 5 See A6 in Appendix A (Standard Appendix). 6.8.3 Electrophoresis support: cellulose acetate membrane. 6.8.4
Electrophoresis conditions: voltage=200 V, time=30 min, migration direction from positive electrode to negative electrode. 6.8.5
Staining method: enzyme color plate method.
Staining solution: See B3 in Appendix B (Standard Appendix). Standard control:
BALB/cJ
C57BL/6J
GB/T 14927.1—2001
6.8.8 Judgment method: Read the band pattern with reference to the above control animals and compare it with Appendix C (Standard Appendix). 6.8.9 Gpi1 electrophoresis result pattern diagram, see Figure 8. b
Gpil electrophoresis result pattern diagram
6.9 Hemoglobin β-chain (H6b) Chr, 76.9.1 Sample: Add 1/4 volume of alkylating agent to hemolysin, see Appendix B (Standard Appendix) B9, 0.3 uL. 6.9.2 Buffer: Tris-Glycine pH8.5 See A6 in Appendix A (Standard Appendix). 6.9.3 Electrophoresis support: Cellulose acetate membrane. 6.9.4 Electrophoresis conditions: Voltage = 200V, time 30min, moving direction from negative electrode to positive electrode. 6.9.5 Staining method: Protein staining method.
6.9.6 Staining solution: See B6 in Appendix B (Standard Appendix). 6.9.7 Standard control,
C57BI./6J
BALB/eJ
6.9.8 Judgment method: Read the band pattern with reference to the above control animals and compare it with Appendix C (Standard Appendix). 6.9.9H66 electrophoresis result pattern diagram, see Figure 9. Hbb
Figure 9 Hbh electrophoresis result pattern
GB/T 14927.1-2001
6. 10 Isocitrate dehydrogenase-1 and malic enzyme-1 (Idh1 and Mex1) Chr.I
Sample: The slurry was diluted with distilled water 1:4, 0.3pL, buffer: Tris-Citrate pH7.6, see Appendix A (Standard Appendix) A3. Electrophoresis conditions: voltage = 200 V, time = 35 min, moving direction from negative electrode to positive electrode. Staining method: Enzyme color plate method, Staining solution: See Appendix B (Standard Appendix) B4 Standard control: Judgment method: Idhl and M. green Result pattern Hasphog 6.11 Phosphoglucose translocation 6.11.1 Sample: kidney plasma, Buffer: 1ris-Glycine monophosphate electrophoresis See Figure! dh2 ModdhI I and Mod1 Electrophoresis Result pattern Figure 18. Appendix A0 Broad electrophoresis Support: Cellulose acetate AG.
Electrophoresis conditions: voltage = 200 V, time threshold = 40 min, moving direction from negative electrode to positive electrode.
Staining solution: See B5 in Appendix B (standard appendix).
6.11.7 Standard control:
C57BL/61
6.11.8 Judgment method: Read the band pattern of the above control animals and compare it with Appendix C (standard appendix).
6.11.9 Pgml electrophoresis result pattern diagram, see Figure 1
6.12 Transferrin (ferransferin, 1r) Chr.96 12.1 Sample: serum, 0.3 μL
6.12.2 Buffer: Tris-Glycine pI18.5 See A6 in Appendix A (standard appendix).
6.12.3 Electrophoresis support: cellulose acetate membrane. 103L、
GB/T 14927.12001
6.5.2 Buffer: Tris-Glycine pII8. 9 See Appendix A (Subject Appendix) A7. 6.5.3 Electrophoresis support: Cellulose acetate membrane, 6.5.4 Electrophoresis conditions: Voltage = 280 V, time 28 min, moving direction from negative electrode to positive electrode. 6.5.5 Staining method: Enzyme color plate method.
6.5.6 Staining solution: See Appendix B (Standard Appendix) B16.5.7 Standard reference:
BALB/eJ
6.5.8 Judgment method: Refer to
6.5.9 ES3 electrophoresis result format
6.6 Esterase-10 (estera
6.6.1 Sample: kidney and liver homogenate
6-6.2 Buffer: Tris-Glye
6.6.3 Ball support: cellulose acetate
slowest band
animal reading
as Figure 5.
10) Chr
C standard
Es3 electrophoresis result pattern
A standard in the appendix
9 grams of nitrogen
high appendix for ratio conversion
6. 6. 4 Electrophoresis conditions: voltage = 280 Time = 28 tons. Moving direction: negative electrode to positive electrode
6.6.5 Staining method: enzyme color plate method
6.6.6 Staining solution: see Appendix B (standard appendix)6.6.7 Standard control:
BALB/eJ
DBA/2J
BUB/BuJ
Slowest band
6.6.8 Judgment method: refer to [the above control animal to read the band type and compare it with Appendix C (standard appendix), 6.6.9Fs10 electrophoresis result pattern diagram, see Figure 66.7 Glucose-t-phosphate dehydrogenase-1 (glucosc.6 phosphatc rlehydrogenase-1.Gpd1) Chr.46.7.1 Sample: fresh kidney or liver homogenate, 0.9l.6.7.2 Buffer: Tris-Glycine plI8.9 See A7 in Appendix A (Standard Appendix). 6.7.3 Electrophoresis support: cellulose acetate plate (membrane). 6.7.4
GB/T 14927. 1: 2001
ES10 electrophoresis result pattern
Electrophoresis conditions: voltage = 200V, time = 50 min, moving direction from negative electrode to positive electrode. Staining method: agar overlay method.
Staining solution: See B7 in Appendix 1 (Standard Appendix). Standard control:
C57BL/6]
BALB/eJ
Slowest band
6.7.8 Judgment method: Read the band pattern of the control animal mentioned above and compare it with Appendix C (Standard Appendix). 6.7.9 Gpal1 electrophoresis result pattern, see Figure? . ■
Figure 7 Gpd1 electrophoresis result pattern
6-8 Glucosephosphateisomerase-1 (Gpil) Chr.76. 8. 1 Sample: hemolysin, 0. 3 μL. .6.8.2
Buffer: Tris-Glycine pH8. 5 See A6 in Appendix A (Standard Appendix). 6.8.3 Electrophoresis support: cellulose acetate membrane. 6.8.4
Electrophoresis conditions: voltage=200 V, time=30 min, migration direction from positive electrode to negative electrode. 6.8.5
Staining method: enzyme color plate method.
Staining solution: See B3 in Appendix B (Standard Appendix). Standard control:
BALB/cJ
C57BL/6J
GB/T 14927.1—2001
6.8.8 Judgment method: Read the band pattern with reference to the above control animals and compare it with Appendix C (Standard Appendix). 6.8.9 Gpi1 electrophoresis result pattern diagram, see Figure 8. b
Gpil electrophoresis result pattern diagram
6.9 Hemoglobin β-chain (H6b) Chr, 76.9.1 Sample: Add 1/4 volume of alkylating agent to hemolysin, see Appendix B (Standard Appendix) B9, 0.3 uL. 6.9.2 Buffer: Tris-Glycine pH8.5 See A6 in Appendix A (Standard Appendix). 6.9.3 Electrophoresis support: Cellulose acetate membrane. 6.9.4 Electrophoresis conditions: Voltage = 200V, time 30min, moving direction from negative electrode to positive electrode. 6.9.5 Staining method: Protein staining method.
6.9.6 Staining solution: See B6 in Appendix B (Standard Appendix). 6.9.7 Standard control,
C57BI./6J
BALB/eJ
6.9.8 Judgment method: Read the band pattern with reference to the above control animals and compare it with Appendix C (Standard Appendix). 6.9.9H66 electrophoresis result pattern diagram, see Figure 9. Hbb
Figure 9 Hbh electrophoresis result pattern
GB/T 14927.1-2001
6. 10 Isocitrate dehydrogenase-1 and malic enzyme-1 (Idh1 and Mex1) Chr.I
Sample: The slurry was diluted with distilled water 1:4, 0.3pL, buffer: Tris-Citrate pH7.6, see Appendix A (Standard Appendix) A3. Electrophoresis conditions: voltage = 200 V, time = 35 min, moving direction from negative electrode to positive electrode. Staining method: Enzyme color plate method, Staining solution: See Appendix B (Standard Appendix) B4 Standard control: Judgment method: Idhl and M. green Result pattern Hasphog 6.11 Phosphoglucose translocation 6.11.1 Sample: kidney plasma, Buffer: 1ris-Glycine monophosphate electrophoresis See Figure! dh2 ModdhI I and Mod1 Electrophoresis Result pattern Figure 18. Appendix A0 Broad electrophoresis Support: Cellulose acetate AG.
Electrophoresis conditions: voltage = 200 V, time threshold = 40 min, moving direction from negative electrode to positive electrode.
Staining solution: See B5 in Appendix B (standard appendix).
6.11.7 Standard control:
C57BL/61
6.11.8 Judgment method: Read the band pattern of the above control animals and compare it with Appendix C (standard appendix).
6.11.9 Pgml electrophoresis result pattern diagram, see Figure 1
6.12 Transferrin (transferrin, 1r) Chr.96 12.1 Sample: serum, 0.3 μL
6.12.2 Buffer: Tris-Glycine pI18.5 See A6 in Appendix A (standard appendix).
6.12.3 Electrophoresis support: cellulose acetate membrane. 103L、
GB/T 14927.12001
6.5.2 Buffer: Tris-Glycine pII8. 9 See Appendix A (Subject Appendix) A7. 6.5.3 Electrophoresis support: Cellulose acetate membrane, 6.5.4 Electrophoresis conditions: Voltage = 280 V, time 28 min, moving direction from negative electrode to positive electrode. 6.5.5 Staining method: Enzyme color plate method.
6.5.6 Staining solution: See Appendix B (Standard Appendix) B16.5.7 Standard reference:
BALB/eJ
6.5.8 Judgment method: Refer to
6.5.9 ES3 electrophoresis result format
6.6 Esterase-10 (estera
6.6.1 Sample: kidney and liver homogenate
6-6.2 Buffer: Tris-Glye
6.6.3 Ball support: cellulose acetate
slowest band
animal reading
as Figure 5.
10) Chr
C standard
Es3 electrophoresis result pattern
A standard in the appendix
9 grams of nitrogen
high appendix for ratio conversion
6. 6. 4 Electrophoresis conditions: voltage = 280 Time = 28 tons. Moving direction: negative electrode to positive electrode
6.6.5 Staining method: enzyme color plate method
6.6.6 Staining solution: see Appendix B (standard appendix)6.6.7 Standard control:
BALB/eJ
DBA/2J
BUB/BuJ
Slowest band
6.6.8 Judgment method: refer to [the above control animal to read the band type and compare it with Appendix C (standard appendix), 6.6.9Fs10 electrophoresis result pattern diagram, see Figure 66.7 Glucose-t-phosphate dehydrogenase-1 (glucosc.6 phosphatc rlehydrogenase-1.Gpd1) Chr.46.7.1 Sample: fresh kidney or liver homogenate, 0.9l.6.7.2 Buffer: Tris-Glycine plI8.9 See A7 in Appendix A (Standard Appendix). 6.7.3 Electrophoresis support: cellulose acetate plate (membrane). 6.7.4
GB/T 14927. 1: 2001
ES10 electrophoresis result pattern
Electrophoresis conditions: voltage = 200V, time = 50 min, moving direction from negative electrode to positive electrode. Staining method: agar overlay method.
Staining solution: See B7 in Appendix 1 (Standard Appendix). Standard control:
C57BL/6]
BALB/eJ
Slowest band
6.7.8 Judgment method: Read the band pattern of the control animal mentioned above and compare it with Appendix C (Standard Appendix). 6.7.9 Gpal1 electrophoresis result pattern, see Figure? . ■
Figure 7 Gpd1 electrophoresis result pattern
6-8 Glucosephosphateisomerase-1 (Gpil) Chr.76. 8. 1 Sample: hemolysin, 0. 3 μL. .6.8.2
Buffer: Tris-Glycine pH8. 5 See A6 in Appendix A (Standard Appendix). 6.8.3 Electrophoresis support: cellulose acetate membrane. 6.8.4
Electrophoresis conditions: voltage=200 V, time=30 min, migration direction from positive electrode to negative electrode. 6.8.5
Staining method: enzyme color plate method.
Staining solution: See B3 in Appendix B (Standard Appendix). Standard control:
BALB/cJ
C57BL/6J
GB/T 14927.1—2001
6.8.8 Judgment method: Read the band pattern with reference to the above control animals and compare it with Appendix C (Standard Appendix). 6.8.9 Gpi1 electrophoresis result pattern diagram, see Figure 8. b
Gpil electrophoresis result pattern diagram
6.9 Hemoglobin β-chain (H6b) Chr, 76.9.1 Sample: Add 1/4 volume of alkylating agent to hemolysin, see Appendix B (Standard Appendix) B9, 0.3 uL. 6.9.2 Buffer: Tris-Glycine pH8.5 See A6 in Appendix A (Standard Appendix). 6.9.3 Electrophoresis support: Cellulose acetate membrane. 6.9.4 Electrophoresis conditions: Voltage = 200V, time 30min, moving direction from negative electrode to positive electrode. 6.9.5 Staining method: Protein staining method.
6.9.6 Staining solution: See B6 in Appendix B (Standard Appendix). 6.9.7 Standard control,
C57BI./6J
BALB/eJ
6.9.8 Judgment method: Read the band pattern with reference to the above control animals and compare it with Appendix C (Standard Appendix). 6.9.9H66 electrophoresis result pattern diagram, see Figure 9. Hbb
Figure 9 Hbh electrophoresis result pattern
GB/T 14927.1-2001
6. 10 Isocitrate dehydrogenase-1 and malic enzyme-1 (Idh1 and Mex1) Chr.I
Sample: The slurry was diluted with distilled water 1:4, 0.3pL, buffer: Tris-Citrate pH7.6, see Appendix A (Standard Appendix) A3. Electrophoresis conditions: voltage = 200 V, time = 35 min, moving direction from negative electrode to positive electrode. Staining method: Enzyme color plate method, Staining solution: See Appendix B (Standard Appendix) B4 Standard control: Judgment method: Idhl and M. green Result pattern Hasphog 6.11 Phosphoglucose translocation 6.11.1 Sample: kidney plasma, Buffer: 1ris-Glycine monophosphate electrophoresis See Figure! dh2 ModdhI I and Mod1 Electrophoresis Result pattern Figure 18. Appendix A0 Broad electrophoresis Support: Cellulose acetate AG.
Electrophoresis conditions: voltage = 200 V, time threshold = 40 min, moving direction from negative electrode to positive electrode.
Staining solution: See B5 in Appendix B (standard appendix).
6.11.7 Standard control:
C57BL/61
6.11.8 Judgment method: Read the band pattern of the above control animals and compare it with Appendix C (standard appendix).
6.11.9 Pgml electrophoresis result pattern diagram, see Figure 1
6.12 Transferrin (ferransferin, 1r) Chr.96 12.1 Sample: serum, 0.3 μL
6.12.2 Buffer: Tris-Glycine pI18.5 See A6 in Appendix A (standard appendix).
6.12.3 Electrophoresis support: cellulose acetate membrane. 10Acetate
Slowest band
Animal reading
Figure 5.
10)Chr
C standard
Es3 electrophoresis result pattern
A standard in the appendix
9 grams of nitrogen
High appendix for ratio conversion
6. 6. 4 Electrophoresis conditions: voltage = 280 Time = 28 tons. Moving direction: negative electrode to positive electrode
6.6.5 Staining method: enzyme color plate method
6.6.6 Staining solution: see Appendix B (standard appendix)6.6.7 Standard control:
BALB/eJ
DBA/2J
BUB/BuJ
Slowest band
6.6.8 Judgment method: refer to [the above control animal to read the band type and compare it with Appendix C (standard appendix), 6.6.9Fs10 electrophoresis result pattern diagram, see Figure 66.7 Glucose-t-phosphate dehydrogenase-1 (glucosc.6 phosphatc rlehydrogenase-1.Gpd1) Chr.46.7.1 Sample: fresh kidney or liver homogenate, 0.9l.6.7.2 Buffer: Tris-Glycine plI8.9 See A7 in Appendix A (Standard Appendix). 6.7.3 Electrophoresis support: cellulose acetate plate (membrane). 6.7.4
GB/T 14927. 1: 2001
ES10 electrophoresis result pattern
Electrophoresis conditions: voltage = 200V, time = 50 min, moving direction from negative electrode to positive electrode. Staining method: agar overlay method.
Staining solution: See B7 in Appendix 1 (Standard Appendix). Standard control:
C57BL/6]
BALB/eJ
Slowest band
6.7.8 Judgment method: Read the band pattern of the control animal mentioned above and compare it with Appendix C (Standard Appendix). 6.7.9 Gpal1 electrophoresis result pattern, see Figure? . ■
Figure 7 Gpd1 electrophoresis result pattern
6-8 Glucosephosphateisomerase-1 (Gpil) Chr.76. 8. 1 Sample: hemolysin, 0. 3 μL. .6.8.2
Buffer: Tris-Glycine pH8. 5 See A6 in Appendix A (Standard Appendix). 6.8.3 Electrophoresis support: cellulose acetate membrane. 6.8.4
Electrophoresis conditions: voltage=200 V, time=30 min, migration direction from positive electrode to negative electrode. 6.8.5
Staining method: enzyme color plate method.
Staining solution: See B3 in Appendix B (Standard Appendix). Standard control:
BALB/cJ
C57BL/6J
GB/T 14927.1—2001
6.8.8 Judgment method: Read the band pattern with reference to the above control animals and compare it with Appendix C (Standard Appendix). 6.8.9 Gpi1 electrophoresis result pattern diagram, see Figure 8. b
Gpil electrophoresis result pattern diagram
6.9 Hemoglobin β-chain (H6b) Chr, 76.9.1 Sample: Add 1/4 volume of alkylating agent to hemolysin, see Appendix B (Standard Appendix) B9, 0.3 uL. 6.9.2 Buffer: Tris-Glycine pH8.5 See A6 in Appendix A (Standard Appendix). 6.9.3 Electrophoresis support: Cellulose acetate membrane. 6.9.4 Electrophoresis conditions: Voltage = 200V, time 30min, moving direction from negative electrode to positive electrode. 6.9.5 Staining method: Protein staining method.
6.9.6 Staining solution: See B6 in Appendix B (Standard Appendix). 6.9.7 Standard control,
C57BI./6J
BALB/eJ
6.9.8 Judgment method: Read the band pattern with reference to the above control animals and compare it with Appendix C (Standard Appendix). 6.9.9H66 electrophoresis result pattern diagram, see Figure 9. Hbb
Figure 9 Hbh electrophoresis result pattern
GB/T 14927.1-2001
6. 10 Isocitrate dehydrogenase-1 and malic enzyme-1 (Idh1 and Mex1) Chr.I
Sample: The slurry was diluted with distilled water 1:4, 0.3pL, buffer: Tris-Citrate pH7.6, see Appendix A (Standard Appendix) A3. Electrophoresis conditions: voltage = 200 V, time = 35 min, moving direction from negative electrode to positive electrode. Staining method: Enzyme color plate method, Staining solution: See Appendix B (Standard Appendix) B4 Standard control: Judgment method: Idhl and M. green Result pattern Hasphog 6.11 Phosphoglucose translocation 6.11.1 Sample: kidney plasma, Buffer: 1ris-Glycine monophosphate electrophoresis See Figure! dh2 ModdhI I and Mod1 Electrophoresis Result pattern Figure 18. Appendix A0 Broad electrophoresis Support: Cellulose acetate AG.
Electrophoresis conditions: voltage = 200 V, time threshold = 40 min, moving direction from negative electrode to positive electrode.
Staining solution: See B5 in Appendix B (standard appendix).
6.11.7 Standard control:
C57BL/61
6.11.8 Judgment method: Read the band pattern of the above control animals and compare it with Appendix C (standard appendix).
6.11.9 Pgml electrophoresis result pattern diagram, see Figure 1
6.12 Transferrin (ferransferin, 1r) Chr.96 12.1 Sample: serum, 0.3 μL
6.12.2 Buffer: Tris-Glycine pI18.5 See A6 in Appendix A (standard appendix).
6.12.3 Electrophoresis support: cellulose acetate membrane. 10Acetate
Slowest band
Animal reading
Figure 5.
10)Chr
C standard
Es3 electrophoresis result pattern
A standard in the appendix
9 grams of nitrogen
High appendix for ratio conversion
6. 6. 4 Electrophoresis conditions: voltage = 280 Time = 28 tons. Moving direction: negative electrode to positive electrode
6.6.5 Staining method: enzyme color plate method
6.6.6 Staining solution: see Appendix B (standard appendix)6.6.7 Standard control:
BALB/eJ
DBA/2J
BUB/BuJ
Slowest band
6.6.8 Judgment method: refer to [the above control animal to read the band type and compare it with Appendix C (standard appendix), 6.6.9Fs10 electrophoresis result pattern diagram, see Figure 66.7 Glucose-t-phosphate dehydrogenase-1 (glucosc.6 phosphatc rlehydrogenase-1.Gpd1) Chr.46.7.1 Sample: fresh kidney or liver homogenate, 0.9l.6.7.2 Buffer: Tris-Glycine plI8.9 See A7 in Appendix A (Standard Appendix). 6.7.3 Electrophoresis support: cellulose acetate plate (membrane). 6.7.4
GB/T 14927. 1: 2001
ES10 electrophoresis result pattern
Electrophoresis conditions: voltage = 200V, time = 50 min, moving direction from negative electrode to positive electrode. Staining method: agar overlay method.
Staining solution: See B7 in Appendix 1 (Standard Appendix). Standard control:
C57BL/6]
BALB/eJ
Slowest band
6.7.8 Judgment method: Read the band pattern of the control animal mentioned above and compare it with Appendix C (Standard Appendix). 6.7.9 Gpal1 electrophoresis result pattern, see Figure? . ■
Figure 7 Gpd1 electrophoresis result pattern
6-8 Glucosephosphateisomerase-1 (Gpil) Chr.76. 8. 1 Sample: hemolysin, 0. 3 μL. .6.8.2
Buffer: Tris-Glycine pH8. 5 See A6 in Appendix A (Standard Appendix). 6.8.3 Electrophoresis support: cellulose acetate membrane. 6.8.4
Electrophoresis conditions: voltage=200 V, time=30 min, migration direction from positive electrode to negative electrode. 6.8.5
Staining method: enzyme color plate method.
Staining solution: See B3 in Appendix B (Standard Appendix). Standard control:
BALB/cJ
C57BL/6J
GB/T 14927.1—2001
6.8.8 Judgment method: Read the band pattern with reference to the above control animals and compare it with Appendix C (Standard Appendix). 6.8.9 Gpi1 electrophoresis result pattern diagram, see Figure 8. b
Gpil electrophoresis result pattern diagram
6.9 Hemoglobin β-chain (H6b) Chr, 76.9.1 Sample: Add 1/4 volume of alkylating agent to hemolysin, see Appendix B (Standard Appendix) B9, 0.3 uL. 6.9.2 Buffer: Tris-Glycine pH8.5 See A6 in Appendix A (Standard Appendix). 6.9.3 Electrophoresis support: Cellulose acetate membrane. 6.9.4 Electrophoresis conditions: Voltage = 200V, time 30min, moving direction from negative electrode to positive electrode. 6.9.5 Staining method: Protein staining method.
6.9.6 Staining solution: See B6 in Appendix B (Standard Appendix). 6.9.7 Standard control,
C57BI./6J
BALB/eJ
6.9.8 Judgment method: Read the band pattern with reference to the above control animals and compare it with Appendix C (Standard Appendix). 6.9.9H66 electrophoresis result pattern diagram, see Figure 9. Hbb
Figure 9 Hbh electrophoresis result pattern
GB/T 14927.1-2001
6. 10 Isocitrate dehydrogenase-1 and malic enzyme-1 (Idh1 and Mex1) Chr.I
Sample: The slurry was diluted with distilled water 1:4, 0.3pL, buffer: Tris-Citrate pH7.6, see Appendix A (Standard Appendix) A3. Electrophoresis conditions: voltage = 200 V, time = 35 min, moving direction from negative electrode to positive electrode. Staining method: Enzyme color plate method, Staining solution: See Appendix B (Standard Appendix) B4 Standard control: Judgment method: Idhl and M. green Result pattern Hasphog 6.11 Phosphoglucose translocation 6.11.1 Sample: kidney plasma, Buffer: 1ris-Glycine monophosphate electrophoresis See Figure! dh2 ModdhI I and Mod1 Electrophoresis Result pattern Figure 18. Appendix A0 Broad electrophoresis Support: Cellulose acetate AG.
Electrophoresis conditions: voltage = 200 V, time threshold = 40 min, moving direction from negative electrode to positive electrode.
Staining solution: See B5 in Appendix B (standard appendix).
6.11.7 Standard control:
C57BL/61
6.11.8 Judgment method: Read the band pattern of the above control animals and compare it with Appendix C (standard appendix).
6.11.9 Pgml electrophoresis result pattern diagram, see Figure 1
6.12 Transferrin (ferransferin, 1r) Chr.96 12.1 Sample: serum, 0.3 μL
6.12.2 Buffer: Tris-Glycine pI18.5 See A6 in Appendix A (standard appendix).
6.12.3 Electrophoresis support: cellulose acetate membrane. 105 See A6 in Appendix A (Standard Appendix). 6.8.3 Electrophoresis support: cellulose acetate membrane. 6.8.4
Electrophoresis conditions: voltage = 200 V, time = 30 min, movement direction from positive to negative. 6.8.5
Staining method: enzyme color plate method.
Staining solution: See B3 in Appendix B (Standard Appendix). Standard control:
BALB/cJ
C57BL/6J
GB/T 14927.1—2001
6. 8. 8 Judgment method: Read the band pattern with reference to the above control animals and compare it with Appendix C (Standard Appendix). 6.8.9 Gpi1 electrophoresis result pattern diagram, see Figure 8. b
Gpil electrophoresis result pattern
6. 9 Hemoglobin β-chain (H6b) Chr, 76. 9. 1 Sample: Add 1/4 volume of alkylating agent to hemolysin, see B9 in Appendix B (Standard Appendix), 0. 3 uL. 6. 9. 2 Buffer: Tris-Glycine pH8. 5 See A6 in Appendix A (Standard Appendix). 6.9.3 Electrophoresis support: Cellulose acetate membrane. 6.9.4 Electrophoresis conditions: Voltage = 200 V, time 30 min, moving direction from negative electrode to positive electrode. 6.9.5 Staining method: protein staining method.
6.9.6 Staining solution: See B6 in Appendix B (Appendix of standard stall). 6.9.7 Standard control,
C57BI./6J
BALB/eJ
6.9.8 Judgment method: Read the band pattern with reference to the above control animals and compare it with Appendix C (Appendix of standard). 6.9.9 H66 electrophoresis result pattern diagram, see Figure 9. Hbb
Figure 9 Hbh electrophoresis result pattern
GB/T 14927.1-2001
6. 10 Isocitrate dehydrogenase-1 and malic enzyme-1 (Idh1 and Mex1) Chr.I
Sample: The slurry was diluted with distilled water 1:4, 0.3pL, buffer: Tris-Citrate pH7.6, see Appendix A (Standard Appendix) A3. Electrophoresis conditions: voltage = 200 V, time = 35 min, moving direction from negative electrode to positive electrode. Staining method: Enzyme color plate method, Staining solution: See Appendix B (Standard Appendix) B4 Standard control: Judgment method: Idhl and M. green Result pattern Hasphog 6.11 Phosphoglucose translocation 6.11.1 Sample: kidney plasma, Buffer: 1ris-Glycine monophosphate electrophoresis See Figure! dh2 ModdhI I and Mod1 Electrophoresis Result pattern Figure 18. Appendix A0 Broad electrophoresis Support: Cellulose acetate AG.
Electrophoresis conditions: voltage = 200 V, time threshold = 40 min, moving direction from negative electrode to positive electrode.
Staining solution: See B5 in Appendix B (standard appendix).
6.11.7 Standard control:
C57BL/61
6.11.8 Judgment method: Read the band pattern of the above control animals and compare it with Appendix C (standard appendix).
6.11.9 Pgml electrophoresis result pattern diagram, see Figure 1
6.12 Transferrin (ferransferin, 1r) Chr.96 12.1 Sample: serum, 0.3 μL
6.12.2 Buffer: Tris-Glycine pI18.5 See A6 in Appendix A (standard appendix).
6.12.3 Electrophoresis support: cellulose acetate membrane. 105 See A6 in Appendix A (Standard Appendix). 6.8.3 Electrophoresis support: cellulose acetate membrane. 6.8.4
Electrophoresis conditions: voltage = 200 V, time = 30 min, movement direction from positive to negative. 6.8.5
Staining method: enzyme color plate method.
Staining solution: See B3 in Appendix B (Standard Appendix). Standard control:
BALB/cJ
C57BL/6J
GB/T 14927.1—2001
6. 8. 8 Judgment method: Read the band pattern with reference to the above control animals and compare it with Appendix C (Standard Appendix). 6.8.9 Gpi1 electrophoresis result pattern diagram, see Figure 8. b
Gpil electrophoresis result pattern
6. 9 Hemoglobin β-chain (H6b) Chr, 76. 9. 1 Sample: Add 1/4 volume of alkylating agent to hemolysin, see B9 in Appendix B (Standard Appendix), 0. 3 uL. 6. 9. 2 Buffer: Tris-Glycine pH8. 5 See A6 in Appendix A (Standard Appendix). 6.9.3 Electrophoresis support: Cellulose acetate membrane. 6.9.4 Electrophoresis conditions: Voltage = 200 V, time 30 min, moving direction from negative electrode to positive electrode. 6.9.5 Staining method: protein staining method.
6.9.6 Staining solution: See B6 in Appendix B (Appendix of standard stall). 6.9.7 Standard control,
C57BI./6J
BALB/eJ
6.9.8 Judgment method: Read the band pattern with reference to the above control animals and compare it with Appendix C (Appendix of standard). 6.9.9 H66 electrophoresis result pattern diagram, see Figure 9. Hbb
Figure 9 Hbh electrophoresis result pattern
GB/T 14927.1-2001
6. 10 Isocitrate dehydrogenase-1 and malic enzyme-1 (Idh1 and Mex1) Chr.I
Sample: The slurry was diluted with distilled water 1:4, 0.3pL, buffer: Tris-Citrate pH7.6, see Appendix A (Standard Appendix) A3. Electrophoresis conditions: voltage = 200 V, time = 35 min, moving direction from negative electrode to positive electrode. Staining method: Enzyme color plate method, Staining solution: See Appendix B (Standard Appendix) B4 Standard control: Judgment method: Idhl and M. green Result pattern Hasphog 6.11 Phosphoglucose translocation 6.11.1 Sample: kidney plasma, Buffer: 1ris-Glycine monophosphate electrophoresis See Figure! dh2 ModdhI I and Mod1 Electrophoresis Result pattern Figure 18. Appendix A0 Broad electrophoresis Support: Cellulose acetate AG.
Electrophoresis conditions: voltage = 200 V, time threshold = 40 min, moving direction from negative electrode to positive electrode.
Staining solution: See B5 in Appendix B (standard appendix).
6.11.7 Standard control:
C57BL/61
6.11.8 Judgment method: Read the band pattern of the above control animals and compare it with Appendix C (standard appendix).
6.11.9 Pgml electrophoresis result pattern diagram, see Figure 1
6.12 Transferrin (ferransferin, 1r) Chr.96 12.1 Sample: serum, 0.3 μL
6.12.2 Buffer: Tris-Glycine pI18.5 See A6 in Appendix A (standard appendix).
6.12.3 Electrophoresis support: cellulose acetate membrane. 10
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