Abcam's Doublestain IHC Kit (ab183285) is designed for use with user supplied rabbit and rat primary antibodies to detect two distinct antigens on human and mouse tissue or cell samples. This kit has been tested on paraffin embedded tissue, however, it can also be used to stain frozen specimens or freshly prepared monolayer cell smears.

Double staining is a common method used in immunohistostaining, allowing for the detection of two distinct antigens in a single tissue. Abcam's Doublestain IHC Kit (ab183285) supplies the user with two polymer enzyme conjugates: Rat HRP Polymer (minimal cross reaction to mouse) and Rabbit AP Polymer with two distinct substrates/chromogens, Emerald and Permanent Red. Emerald chromogen reacts with the Rat HRP polymer conjugate to produce a green color. Permanent Red reacts with the Rabbit AP polymer to produce a red color. When two proteins are co-localized a blue/ purple color will develop depending on which antigen is stronger. If only the rat antigen is present only the Emerald chromogen will be present and if the rabbit antigen is present only the Permanent Red chromogen will be present. Abcam's Doublestain IHC Kit (ab183285) is a non-biotin system avoiding the extra steps involved in blocking non-specific binding due to endogenous biotin.

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• Multicolor chromogenic