Biotinylated Goat Anti-Rabbit IgG (H+L) (Ready to use)

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The ab64256 reagent is used in a labeled streptavidin biotin immunoenzymatic antigen detection system. This technique involves the sequential incubation of the specimen with an unconjugated primary antibody specific to the target antigen, a biotinylated secondary antibody which reacts with the primary antibody, enzyme labeled streptavidin, and substrate chromogen.

Staining protocol

• Deparaffinize and rehydrate formalin-fixed paraffin-embedded tissue section.

• Add appropriate hydrogen peroxide blocking solution to cover the sections. Incubate for 10 minutes. Wash 2 times in buffer.

• Perform appropriate pretreatment if required. Wash 3 times in buffer.

• Apply appropriate protein blocking solution and incubate for 5 minutes at room temperature to block non specific background staining. Wash 1 time in buffer.

• Apply primary antibody and incubate according to manufacturer's protocol.

• Wash 4 times in buffer. Apply Ready to use Biotinylated Goat anti Rabbit IgG (ab64256) solution and incubate for 10-15 minutes at room temperature. Wash 4 times in buffer.

• Apply enzyme labelled streptavidin and incubate according to manufacturer's protocol.

• Rinse 4 times in buffer. Place slide in appropriate chromogenic substrate and incubate until desired reaction is achieved. Rinse 4 times in buffer.

• Add enough drops of Haematoxylin to cover the section. Incubate for 1 minute.

• Rinse 7-8 times in tap water. Add Mounting Medium to cover the section.
  
  Find complete IHC kits, and reagents for antigen retrieval, blocking, signal amplification, visualization, counterstaining, and mounting in the IHC kits and reagents guide.