Immunoblotting: Antigen Detection Using Chemiluminescence-SUBSCRIPTION REQUIRED Protocol

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Protocol Information

Title Immunoblotting: Antigen Detection Using Chemiluminescence-SUBSCRIPTION REQUIRED
Source CSH Protocols; 2006; doi:10.1101/pdb.prot4271

Overview

The blot is blocked to prevent nonspecific adsorption of the immunological reagents. Antibodies are then bound to the proteins immobilized on the membrane, and the antigen is detected by labeling the antibodies with conveniently identified tags. Common labeling methods for chemiluminescent detection include anti-immunoglobulin antibody-coupled enzymes such as horseradish peroxidase, which catalyzes the oxidation of luminol and in turn releases light. This light emission can be captured on standard X-ray film to yield an excellent record that is easy to display; however, it is difficult to establish accurate quantitation of protein levels, and light emission cannot be easily controlled

Details

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