Directly conjugated antibodies for immunofluorescence

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Pre-adsorbed secondary antibodies

Minimize non-specific binding and high background staining.

Great secondary antibody. Good specificity and strong signal. Allows a big dilution and is still sensitive enough. I would recommend this antibody to anyone.

- Researcher from University of Cambridge, United Kingdom

Pre-adsorption (also referred to as cross-adsorption) is an extra purification step introduced to increase the specificity of an 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. The result of the procedure is a secondary antibody that recognizes rabbit IgG light chains with high specificity.

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.

Abcam’s pre-adsorbed secondary antibodies

Regardless of the immunoassay or sample you use, we offer a broad range of high quality pre-adsorbed secondary antibodies to satisfy your needs:

  • Against more than 15 different species and their isotypes including mouse, rabbit, rat, goat, and chicken.
  • Raised in different species such as goat and donkey.
  • Unconjugated and conjugated to diverse fluorescent dyes (e.g. Alexa Fluor®), enzymes (e.g. HRP), and proteins (e.g. biotin).
  • Both whole-molecule and fragment secondary antibodies are available.
  • Validated in diverse applications including western blot (WB), immunofluorescence (ICC/IF), immunohistochemistry (IHC), ELISA, and flow cytometry.
  • Cited in more than 270 peer-reviewed publications with a 4.5 stars average customer rating.

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

Product NameApplicationsab ID
Anti-mouse Alexa Fluor® 405 (IgG H&L)ICC/IF, ELISA, IHC-P, IHC-Fr, Flow Cytab175661
Anti-rabbit Alexa Fluor® 405 (IgG H&L)ICC/IF, ELISA, IHC-P, IHC-Fr, Flow Cytab175648
Anti-rat Alexa Fluor® 405 (IgG H&L)ICC/IF, ELISA, IHC-P, IHC-Fr, Flow Cytab175670
Anti-goat Alexa Fluor® 405 (IgG H&L)ICC/IF, ELISA, IHC-P, IHC-Fr, Flow Cytab175665
Anti-chicken Alexa Fluor® 405 (IgY H&L)ICC/IF, ELISA, IHC-P, IHC-Fr, Flow Cytab175675

Preadsorbed Alexa Fluor® 488 conjugated secondary antibodies

Product NameApplicationsab ID
Anti-mouse Alexa Fluor® 488 (IgG H&L)ICC/IF, ELISA, IHC-P, IHC-Fr, Flow Cytab150117
Anti-rabbit Alexa Fluor® 488 (IgG H&L)ICC/IF, ELISA, IHC-P, IHC-Fr, Flow Cytab150081
Anti-rat Alexa Fluor® 488 (IgG H&L)ICC/IF, ELISA, IHC-P, IHC-Fr, Flow Cytab150153
Anti-goat Alexa Fluor® 488 (IgG H&L)ICC/IF, ELISA, IHC-P, IHC-Fr, Flow Cytab150133
Anti-chicken Alexa Fluor® 488 (IgY H&L)ICC/IF, ELISA, IHC-P, IHC-Fr, Flow Cytab150173

Preadsorbed Alexa Fluor® 555 conjugated secondary antibodies

Product NameApplicationsab ID
Anti-mouse Alexa Fluor® 555 (IgG H&L)ICC/IF, ELISA, IHC-P, IHC-Fr, Flow Cytab150110
Anti-rabbit Alexa Fluor® 555 (IgG H&L)ICC/IF, ELISA, IHC-P, IHC-Fr, Flow Cytab150086
Anti-rat Alexa Fluor® 555 (IgG H&L)ICC/IF, ELISA, IHC-P, IHC-Fr, Flow Cytab150154
Anti-goat Alexa Fluor® 555 (IgG H&L)ICC/IF, ELISA, IHC-P, IHC-Fr, Flow Cytab150134
Anti-chicken Alexa Fluor® 555 (IgY H&L)ICC/IF, ELISA, IHC-P, IHC-Fr, Flow Cytab150174

Preadsorbed Alexa Fluor® 594 conjugated secondary antibodies

Product NameApplicationsab ID
Anti-mouse Alexa Fluor® 594 (IgG H&L)ICC/IF, ELISA, IHC-P, IHC-Fr, Flow Cytab150120
Anti-rabbit Alexa Fluor® 594 (IgG H&L)ICC/IF, ELISA, IHC-P, IHC-Fr, Flow Cytab150064
Anti-rat Alexa Fluor® 594 (IgG H&L)ICC/IF, ELISA, IHC-P, IHC-Fr, Flow Cytab150156
Anti-goat Alexa Fluor® 594 (IgG H&L)ICC/IF, ELISA, IHC-P, IHC-Fr, Flow Cytab150136
Anti-chicken Alexa Fluor® 594 (IgY H&L)ICC/IF, ELISA, IHC-P, IHC-Fr, Flow Cytab150176

Preadsorbed Alexa Fluor® 647 conjugated secondary antibodies

Product NameApplicationsab ID
Anti-mouse Alexa Fluor® 647 (IgG H&L)ICC/IF, ELISA, IHC-P, IHC-Fr, Flow Cytab150119
Anti-rabbit Alexa Fluor® 647 (IgG H&L)ICC/IF, ELISA, IHC-P, IHC-Fr, Flow Cytab150083
Anti-rat Alexa Fluor® 647 (IgG H&L)ICC/IF, ELISA, IHC-P, IHC-Fr, Flow Cytab150155
Anti-goat Alexa Fluor® 647 (IgG H&L)ICC/IF, ELISA, IHC-P, IHC-Fr, Flow Cytab150135
Anti-chicken Alexa Fluor® 647 (IgY H&L)ICC/IF, ELISA, IHC-P, IHC-Fr, Flow Cytab150175

Pre-adsorbed HRP conjugated secondary antibodies

Product NameApplicationsab ID
Anti-mouse HRP (IgG H&L)IHC-P, ELISA, WBab97040
Anti-rabbit HRP (IgG H&L)ICC, IHC-P, ELISA, WBab97080
Anti-rat HRP (IgG H&L)ICC/IF, IHC-P, IHC-Fr, WB, ELISA, Dot Blotab7097
Anti-goat HRP (IgG H&L)ICC, IHC-P, ELISA, WBab97120
Anti-chicken HRP (IgY H&L)WB, Immunomicroscopy, ICC/IF, Dot Blot, ELISA, IHC-P, IHC-Frab7118

Pre-adsorbed biotinylated secondary antibodies

Product NameApplicationsab ID
Anti-mouse Biotin (IgG H&L)ICC/IF, Dot Blot, ELISA, IHC-P, IHC-Fr, Immunomicroscopy, WBab7074
Anti-rabbit Biotin (IgG H&L)ICC/IF, WB, ELISA, Immunomicroscopy, Dot Blot, IHC-P, IHC-Frab7082
Anti-rat Biotin (IgG H&L)ICC/IF, WB, ELISA, Immunomicroscopy, Dot Blot, IHC-P, IHC-Frab7096
Anti-goat Biotin (IgG H&L)ICC/IF, Dot Blot, ELISA, IHC-P, IHC-Fr, Immunomicroscopy, WBab7124
Anti-chicken Biotin (IgY H&L)ICC/IF, Dot Blot, ELISA, IHC-P, IHC-Fr, Immunomicroscopy, WBab7117

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