The DoubleStain IHC kit (ab183281) is designed to be used with user supplied goat and rat primary antibodies to detect two distinct antigens on human or mouse tissue or cell samples. The kit has been tested on paraffin embedded human and mouse tissues, however, it can also be used on frozen specimen and freshly prepared monolayer cell smears.

Double staining is one of the most common methods used in immunohistostaining allowing the detection of two distinct antigens in a single tissue. Abcam's DoubleStain IHC Kit (ab183281) supplies the user with two polymer enzyme conjugates: HRP polymer anti-Goat IgG and AP polymer anti-Rat IgG with two distinct substrates/chromogens: BCIP/NBT (Purple) and AEC (Red). Users sequentially apply the two enzyme conjugates onto the specimen. When two proteins are present a purple/red color will develop depending on the presence and location of the antigens the two colors should be distinct. If only the anti-goat antigen is present only Red will be visible and if the anti-Rat antigen is present Purple will be present. The kit is non-biotin system avoiding endogenous biotin non-specific binding.

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