Western Blot is a widely used method for detecting target proteins. This method uses polyacrylamide gel electrophoresis to separate the total protein sample according to the molecular weight, and then transfers the protein on the gel to the PVDF membrane or NC membrane of the solid carrier through electroporation. The specific antibody (primary antibody) reacts with the target protein, and then the secondary antibody with HRP enzyme tag combines with the primary antibody. The color reagent reacts with the enzyme labeled secondary antibody to determine the expression of the target protein in cells or tissues according to the color location and signal strength. Steps of protein immunoblotting experiment

Attention: This experimental procedure is a routine Western Blot experiment. As WB experiments have been improved in different laboratories, the experimental conditions are different, and the experimental methods and formulas are not completely consistent, it is necessary to combine specific experiments for reasonable operation. This protocol is from a research institution and is for reference only.

1、 Preparation of experimental materials and reagents