Polypeptide inhibiting porcine PD-1/PD-L1 pathway and application thereof
A PD-L1, PD-1 technology, applied in medical preparations containing active ingredients, specific peptides, peptide/protein components, etc., can solve problems such as increased expression and T cell failure
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[0022] The present invention will be described in detail below in conjunction with the embodiments.
[0023] Peptide-labeled FITC
[0024] Use the FITC Antibody Labeling Kit to label FITC on the polypeptide. The FITC Antibody Labeling Kit can connect FITC to amino acid residues such as sulfhydryl, imidazole, carbonyl, and tyrosyl. The steps are as follows:
[0025] (1) Peptide preparation, add 40 μL of 0.67 mol / L sodium borate solution to 0.5 mL of 2 mg / mL peptide solution, and adjust the peptide concentration to 1 mg / mL;
[0026] (2) Add 0.5mL polypeptide into a tube containing about 30μL FITC reagent, mix well to fully dissolve the reagent, and incubate at room temperature in the dark for 60min;
[0027] (3) Add 400 μL of purified resin to the reaction tube, put the reaction tube into a micro collection tube, centrifuge at room temperature for 30-45 sec at 1000 r / min, and discard the collection tube;
[0028] (4) Put the reaction tube into a new collection tube, add 250-27...
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