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Reviewed July 6, 2021
Pre-adsorption (also referred to as cross-adsorption) is an extra purification step introduced to increase the specificity of a secondary antibody. The solution containing secondary antibodies is passed through a column matrix containing immobilized serum proteins from potentially cross reactive species. Non-specific secondary antibodies are retained in the column, while highly specific secondaries flow through.
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.
We always recommend using pre-adsorbed (cross-absorbed) secondary antibodies to minimize non-specific background when working with tissues and cells. This means pre-adsorbed antibodies are useful in many different experiments, however they are most useful for:
Example: Use an anti-rabbit Alexa Fluor® 488 pre-adsorbed secondary against mouse serum and an anti-mouse Alexa Fluor® 647 pre-adsorbed secondary against rabbit serum when performing a double staining experiment using primary antibodies raised in rabbit and mouse respectively.
Example: Use an anti-mouse HRP pre-adsorbed secondary antibody against human serum when staining human tissues or cell lines.
All pre-adsorbed Alexa Fluor® secondary antibodies listed below are suitable for use in ICC/IF, ELISA, IHC-P, IHC-Fr and flow cytometry.
Please note some of our pre-adsorbed HRP secondary antibodies are only suitable for use in specific applications, these are listed below.
Product Name | Applications |
Anti-mouse HRP (IgG H&L) (ab97040) | IHC-P, ELISA, WB |
Anti-rabbit HRP (IgG H&L) (ab97080) | ICC, IHC-P, ELISA, WB |
Anti-rat HRP (IgG H&L) (ab7097) | ICC/IF, IHC-P, IHC-Fr, WB, ELISA, Dot Blot |
Anti-goat HRP (IgG H&L) (ab97120) | ICC, IHC-P, ELISA, WB |
Anti-chicken HRP (IgY H&L) (ab7118) | WB, Immunomicroscopy, ICC/IF, Dot Blot, ELISA, IHC-P, IHC-Fr |
Please note some of our pre-adsorbed biotinylated secondary antibodies are only suitable for use in specific applications, these are listed below
Product Name | Applications |
Anti-mouse Biotin (IgG H&L) (ab7074) | ICC/IF, Dot Blot, ELISA, IHC-P, IHC-Fr, Immunomicroscopy, WB |
Anti-rabbit Biotin (IgG H&L) (ab7082) | ICC/IF, WB, ELISA, Immunomicroscopy, Dot Blot, IHC-P, IHC-Fr |
Anti-rat Biotin (IgG H&L) (ab7096) | ICC/IF, WB, ELISA, Immunomicroscopy, Dot Blot, IHC-P, IHC-Fr |
Anti-goat Biotin (IgG H&L) (ab7124) | ICC/IF, Dot Blot, ELISA, IHC-P, IHC-Fr, Immunomicroscopy, WB |
Anti-chicken Biotin (IgY H&L) (ab7117) | ICC/IF, Dot Blot, ELISA, IHC-P, IHC-Fr, Immunomicroscopy, WB |
Alexa Fluor® is a registered trademark of Life Technologies. Alexa Fluor® dye conjugates contain(s) technology licensed to Abcam by Life Technologies.