DoubleStain IHC Kit: R&Rt on human/mouse tissue (Green/HRP & AP/Red) (ab183285)


  • Product name
    DoubleStain IHC Kit: R&Rt on human/mouse tissue (Green/HRP & AP/Red)
  • Assay type
    Polymer Multiplex
  • Product overview

    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.

  • Tested applications
    Suitable for: IHC-Fr, IHC-Pmore details


  • Storage instructions
    Store at +4°C. Please refer to protocols.
  • Components Identifier 12 ml 36 ml 120 ml
    Emerald Chromogen 1 x 15ml 2 x 18ml 1 x 60ml
    Non-aqueous Mounting Medium 1 x 12ml 2 x 18ml
    Permanent Red Activator (5x) 1 x 3ml 1 x 7.2ml 2 x 12ml
    Permanent Red Chromogen (100x) 1 x 150µl 1 x 360µl 1 x 1.2ml
    Permanent Red Substrate 1 x 15ml 2 x 18ml 1 x 120ml
    Rabbit AP Polymer 1 x 6ml 1 x 18ml 1 x 60ml
    Rat HRP Polymer 1 x 6ml 1 x 18ml 1 x 60ml
  • Research areas


Our Abpromise guarantee covers the use of ab183285 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-Fr Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.



ab183285 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

IHC-P on mouse liver tissue for Il-18 and F4/80

Excellent Excellent 5/5 (Ease of Use)
DoubleStain IHC Kit: R&Rt on human/mouse tissue (Green/HRP & AP/Red) was used on parraffin-embedded mouse liver sections with tumors to overcome autofluorescence problem. After de-waxing with xylene and antigen retrieval in citrate buffer, I immunolabeled the tissue with Il-18 and F4/80 . I followed the user's recommendations and I observed very little or mild background with the Permanent Red and almost no background with the Emerald staining. I preferred not to counterstain the tissue section with hematoxylin, as it was easier to see the dark blue codistribution wihout it.
Figure legend
Immunohistochemical co-distribution of F4/80 and Il18 expressing cells in mouse liver with colorectal cancer metastatic growth. Staining of WT or IL18-/- mice.

Ms. Claudia Champagne

Verified customer

Submitted Jun 30 2015

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