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Pre-adsorbed secondary antibodies
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.
Advantages of pre-adsorbed secondary antibodies
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:
- Multicolor experiments: When several primaries 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.
- 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.
- 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 an anti-mouse HRP pre-adsorbed secondary antibody against human serum when staining human tissues or cell lines.
Pre-adsorbed Alexa Fluor® conjugated secondary antibodies
All pre-adsorbed Alexa Fluor® secondary antibodies listed below are suitable for ICC/IF, ELISA, IHC-P, IHC-Fr and flow cytometry.
| Conjugate | Product name |
| Alexa Fluor® 405 | |
Anti-rabbit Alexa Fluor® 405 (IgG H&L) preadsorbed (ab175648) | |
Anti-rat Alexa Fluor® 405(IgG H&L) preadsorbed (ab175670) | |
Anti-goat Alexa Fluor® 405 (IgG H&L) preadsorbed (ab175665) | |
Anti-chicken Alexa Fluor® 405(IgY H&L) preadsorbed (ab175675) | |
Alexa Fluor® 488 | Anti-mouse Alexa Fluor® 488 (IgG H&L) preadsorbed (ab150117) |
| Anti-rabbit Alexa Fluor® 488 (IgG H&L) preadsorbed (ab150081) | |
| Anti-rat Alexa Fluor® 488 (IgG H&L) preadsorbed (ab150153) | |
| Anti-goat Alexa Fluor® 488 (IgG H&L) preadsorbed (ab150133) | |
| Anti-chicken Alexa Fluor® 488 (IgY H&L) preadsorbed (ab150173) | |
| Alexa Fluor® 555 | Anti-mouse Alexa Fluor® 555 (IgG H&L) (ab150110) |
| Anti-rabbit Alexa Fluor® 555 (IgG H&L) (ab150086) | |
| Anti-rat Alexa Fluor® 555 (IgG H&L) (ab150154) | |
| Anti-goat Alexa Fluor® 555 (IgG H&L) (ab150134) | |
| Anti-chicken Alexa Fluor® 555 (IgY H&L) (ab150174) | |
| Alexa Fluor® 594 | Anti-mouse Alexa Fluor® 594 (IgG H&L) (ab150120) |
| Anti-rabbit Alexa Fluor® 594 (IgG H&L) (ab150064) | |
| Anti-rat Alexa Fluor® 594 (IgG H&L) (ab150156) | |
| Anti-goat Alexa Fluor® 594 (IgG H&L) (ab150136) | |
| Anti-chicken Alexa Fluor® 594 (IgY H&L) (ab150176) | |
| Alexa Fluor® 647 | Anti-mouse Alexa Fluor® 647 (IgG H&L) (ab150119) |
| Anti-rabbit Alexa Fluor® 647 (IgG H&L) (ab150083) | |
| Anti-rat Alexa Fluor® 647 (IgG H&L) (ab150155) | |
| Anti-goat Alexa Fluor® 647 (IgG H&L) (ab150135) | |
| Anti-chicken Alexa Fluor® 647 (IgY H&L) (ab150175) |
Pre-adsorbed HRP conjugated secondary antibodies
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 |
Pre-adsorbed biotinylated secondary antibodies
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.