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- 2025年07月09日
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文献和实验of interest available; 2) formation of the antibody-antigen complex by adding specific antibody; 3) separation and detection of the immune complex by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting. Both polyclonal and monoclonal
PROTOCOL To 1.5 mL eppendorf tubes add: 200 µg of protein extract (see Western blot protocol for protein sample preps) q.s. to 300 µL with RIPA (with protease and phosphatase inhibitors). Primary antibody (amount determined
CO-IMMUNOPRECIPITATION ANALYSIS OF PROTEIN-PROTEIN INTERACTIONS
into a 15 ml tube. For immunoprecipitation, add 10 ml (polyclonal serum) or 10 mgs (monoclonal antibody) to 1 ml of extract. Incubate with end-over-end mixing for 2 hours at 40 C. Save 50 ml of each cell extract and add 50 ml of 2X SDS-PAGE sample buffer
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