When a mouse monoclonal antibody is desired for immunohistochemical detection on murine tissues, the biotinylated secondary antibody used for detection will bind to the primary antibody and will bind to endogenous mouse IgG in the tissue. Troublesome background staining may occur from bound mouse IgG.
The majority of primary antibodies on the market are not biotinylated. It is very expensive and time consuming to biotinylate an antibody. Usually, a minimum 2-5 mg of antibody is necessary for the procedure, and the total amount (mg/ml) of the antibody is reduced through the process.
To circumvent the problem, Biocare has developed the MM Biotinylation Kit. The kit provides all the reagents necessary to biotinylate a primary antibody.
The basic concept of the kit is very simple. Prior to the application of primary antibody, the antibody is mixed with a biotinylation reagent for 30 minutes. As little as 5 to 10 μl of antibody can be used. Once the complex is formed, it can be immediately applied to the tissue specimen. The tissue is then incubated with streptavidin-peroxidase, and the color-reaction is developed by a diaminobenzidine/hydrogen peroxide chromogen-substrate (DAB).
The system has been specially formulated to eliminate endogenous mouse IgG background staining. Even the most difficult tissues such as lymphatic, lung and kidney tissues are virtually background free.