Abcam's Doublestain IHC Kit (ab183282) 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.

Double staining is one of most common methods used in immunohistostaining allowing for the detection of two distinct antigens in a single tissue. Abcam's Doublestain IHC Kit (ab183282) supplies the user with two polymer enzyme conjugates: Goat AP Polymer and Rat HRP Polymer with two distinct substrates/chromogens, Emerald and Permanent Red. With this protocol the goat and rat primary antibodies will be applied to the tissue at the same time but the user will sequentially apply the polymer enzyme conjugate to the specimen. When two proteins are present in the same location a blue or purple color will develop depending on the quantity of each antigen screened. If only the anti-goat antigen is present only the Permanent Red chromogen will be visible and if only the Rat antigen is present then only the Emerald chromogen will be seen. Abcam's Doublestain IHC Kit (ab183282) is a non-biotin system avoiding endogenous biotin non-specific binding.

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• Multicolor chromogenic