Secondary Antibody Protocols and Resources

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Below you will find links to a selection of protocols and useful resources for various applications using secondary antibodies. For our full set of resources please go to our antibody resources page.

Immunohistochemistry

  • Protocol – general immunostaining protocol, with links to other protocols
  • Tips when using a secondary antibody in IHC
IHC -  Mouse spleen cryosection stained with Rat Anti-Mouse F4/80 Antibody, clone Cl:A3-1 (MCA497G)

Mouse spleen cryosection stained with Rat Anti-Mouse F4/80 Antibody, clone Cl:A3-1 (MCA497G) at a 1/100 dilution followed by Goat Anti-Rat IgG:HRP (STAR72) at a 1/50 dilution.


Western Blotting

  • Protocol – general western blotting protocols
  • Fluorescent antibodies – StarBright™ Blue 700 Antibodies (targeting mouse and rabbits) are ideal for the detection of single target proteins and multiplex detection of several proteins
Western Blotting - Western blot analysis of HEK293 embryonic kidney cell line whole cell lysate probed with Mouse Anti- Human H-Ras Antibody (MCA2884)

Western blot analysis of HEK293 embryonic kidney cell line whole cell lysate probed with Mouse Anti-Human H-Ras Antibody (MCA2884) followed by HRP conjugated Goat Anti-Mouse IgG Secondary Antibody (STAR117P) visualized using chemiluminescence.


ELISA

  • Protocol – general indirect ELISA protocol, with links to competitive ELISA protocol
  • Tips for ELISA, including antibody considerations
ELISA - Sandwich ELISA analysis of human IgE using Mouse Anti-Human IgE Antibody (0100-0413)

Sandwich ELISA analysis of human IgE using Mouse Anti-Human IgE Antibody (0100-0413) as a capture reagent and biotinylated Goat Anti-Human IgE Antibody (STAR147B), as a detection reagent, with recombinant human IgE (HCA171) as antigen for generation of the standard curve. Detection is by HRP conjugated streptavidin and substrate. Microtitre plate is read at O.D. 450 nm on the Bio-Rad iMark™ Microplate Absorbance Reader (1681135). Plasma and serum samples diluted 1:5 are indicated in green and orange respectively.


Flow Cytometry

  • Protocol – general indirect immunofluorescence staining of cells and blood protocol
  • Products, protocols, tips and tricks for all your flow cytometry needs
Flow Cytometry - RPE conjugated Rat Anti-Dog CD4 (MCA1038PE) and Mouse IgG1 Isotype Control (MCA928) labeled with Goat Anti-Mouse FITC (STAR117F).

A, RPE conjugated Rat Anti-Dog CD4 (MCA1038PE) and Mouse IgG1 Isotype Control (MCA928) labeled with Goat Anti-Mouse FITC (STAR117F). B, RPE conjugated Rat Anti-Dog CD4 (MCA1038PE) and Mouse Anti-Dog CD3 (MCA1774GA) labeled with Goat Anti-Mouse FITC (STAR117F). All experiments performed on red cell lysed canine blood gated lymphoid cells in the presence of 10% dog serum. Data acquired on the ZE5™ Cell Analyzer.


Browse application specific web pages to quickly and easily find antibodies, kits and reagents tested in your application of choice, and access many scientific tools such as protocols, webinars, tips and in-depth guides, all in one place.


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