Porcine P21 protein antibody as well as preparation method and application thereof
A P21, protein technology, applied in the direction of serum immunoglobulin, anti-animal/human immunoglobulin, material test products, etc. Strong demand for antibodies and other issues, to achieve the effect of broad application prospects
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Embodiment 1
[0033] The preparation of pig p21 protein antibody, the technical route is as follows figure 1 , Including the following steps:
[0034] S1. Cloning of pig P21 gene
[0035] In the present invention, the total RNA of porcine skeletal muscle satellite cells is first extracted, cDNA is obtained by reverse transcription, and the porcine P21 gene is cloned by using the cDNA as a template. It includes the following steps:
[0036] S11. Extraction of total RNA: For extraction of total RNA from porcine skeletal muscle satellite cells, refer to the instruction manual of RNAiso Plus Total RNA Extraction Reagent from TAKARA.
[0037] S12. Reverse transcription of total RNA: The reverse transcription of the extracted total RNA is performed according to the instructions of TIANGEN's TIANScript cDNA first-strand synthesis kit. The specific steps are as follows:
[0038] Add the following reaction mixture to a nuclease-free centrifuge tube in an ice bath:
[0039]
[0040] Reaction conditions: After ...
Embodiment 2
[0118] The specificity of the anti-pig P21 antibody prepared in Example 1 of the present invention is compared with the commercially available anti-mouse P21 antibody. The specific method is: comparing the anti-pig P21 antibody prepared by the present invention with the commercially available anti-mouse P21 antibody by Western blot. The ability of P21 antibody to recognize P21 protein of pigs and mice, such as Picture 11 Shown from Picture 11 It can be seen that the anti-pig P21 antibody prepared in Example 1 of the present invention can well recognize the P21 protein of pigs and mice; while the anti-mouse P21 antibody purchased on the market can only recognize the P21 protein of mice to a limited extent. The P21 protein of pigs is basically not recognized. It can be seen that the anti-pig P21 protein antibody prepared by the present invention has specificity to the pig P21 protein.
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