Pre-adsorbed secondary antibodies

^{Figure 1. In this example, a solution of secondary antibodies recognizing rabbit IgG light chains is passed through a matrix containing immobilized serum proteins from potentially cross-reactive species such as sheep and bovine IgGs. Only antibodies highly specific to rabbit IgG light chain will flow through the column, while secondary antibodies cross-reacting with sheep or bovine IgG light chains will remain bound to the proteins in the column. The result of the procedure is a secondary antibody that recognizes rabbit IgG light chains with high specificity.}

Advantages of using pre-adsorbed (cross-adsorbed) secondary antibodies

We always recommend using pre-adsorbed (cross-adsorbed) secondary antibodies to minimize non-specific background when working with tissues and cells. However, these antibodies are specifically useful in:

• Multicolor experiments: When several primary and their corresponding secondary antibodies are used simultaneously, the secondary antibody should be pre-adsorbed against the species in which the other primary antibodies have been raised to reduce the risk of cross reactivity between them.

Example: Use secondary antibodies anti-rabbit Alexa Fluor® 488 pre-adsorbed against mouse serum and an anti-mouse Alexa Fluor® 647 pre-adsorbed against rabbit serum when performing a double staining experiment using primary antibodies raised in rabbit and mouse respectively.

• IHC experiments on samples with abundant amounts of endogenous immunoglobulins (Igs): The secondary antibody should be pre-adsorbed against the same species the sample originated from to reduce the risk of cross reactivity between the secondary antibody and endogenous Igs.

Example: Use secondary antibody anti-mouse HRP pre-adsorbed against human serum when staining human tissues or cell lines.

Below you will find links to our most popular pre-adsorbed secondary antibodies. If you would like to explore our broad offering of pre-adsorbed secondaries, you can also use our search engine here:

Preadsorbed Alexa Fluor® 405 conjugated secondary antibodies

Alexa Fluor® is a registered trademark of Life Technologies. Alexa Fluor® dye conjugates contain(s) technology licensed to Abcam by Life Technologies.