Swine GAPDH (Glyceraldehyde-3-Phosphate Dehydrogenase) protein antibody and preparation method and application thereof

[0034] S1. Cloning of porcine GAPDH gene: In the present invention, firstly, total RNA of porcine skeletal muscle satellite cells was extracted, reverse-transcribed to obtain cDNA, and porcine GAPDH gene was cloned using this as a template.

[0035] S11. For the extraction of total RNA from porcine skeletal muscle satellite cells, refer to the instruction manual of RNAiso Plus Total RNA Extraction Reagent from TAKARA Company.

[0036] S12. Reverse transcription of total RNA: The reverse transcription of RNA was performed according to the instructions of the TIANScript cDNA first-strand synthesis kit from TIANGEN, and the specific steps were as follows:

[0037] Add the following reaction mixture to a nuclease-free centrifuge tube on ice:

[0038]

[0039] Reaction conditions: heat at 70°C for 5 minutes, then quickly cool on ice for 2 minutes, briefly centrifuge to collect the reaction solution, and then add the following components:

[0040]

[0041] Reaction condition...

[0115] The specificity of the anti-pig GAPDH antibody prepared in Example 1 of the present invention was compared with the commercially available anti-human GAPDH antibody. The specific method was: compare the anti-pig GAPDH antibody prepared by the present invention with the commercially available anti-human GAPDH antibody by Western blot As can be seen from Figure (14), the anti-pig GAPDH antibody prepared in Example 1 of the present invention can well recognize the GAPDH protein of pigs and mice; Human GAPDH antibody can only limitedly recognize mouse GAPDH protein, and basically cannot recognize pig GAPDH protein.