Overview

  • Product name

    TripleStain IHC Kit: R&R&M on rodent tissue (DAB, AP/Red & DAB/Ni)
  • Assay type

    Polymer Multiplex
  • Product overview

    The TripleStain IHC Kit (ab183296) is designed to use with user supplied one mouse and two rabbit primary antibodies to detect three distinct antigens on a single mouse/rat tissue or cell samples. The kit has been tested on tissue specimens that are paraffin embedded. For frozen tissue a lower temperature of 65ºC must be used for Antibody Blocker to prevent tissue from dissociating from slide. 

     

    Triple staining is a common method used in immunohistochemistry for the detection of three distinct antigens and their co-expression in a single tissue. Abcam’s TripleStain IHC Kit supplies polymer enzyme conjugates: polymer-HRP anti-mouse IgG, polymer-HRP anti-rabbit IgG, and polymer-AP anti-rabbit IgG with three substrates/chromogens; DAB (brown), DAB-Ni (black), and Permanent Red (Red). The TripleStain Kit is a non-biotin system avoiding non-specific background due to endogenous biotin binding. This kit has been optimized to have no cross detection when detecting two primary antibodies from the same host species using a unique blocking system. Simplified steps allow users to complete triple staining within 5 hours (without antigen retrieval) or 6 hours (with antigen retrieval). The well tested protocol provides users with the ability to permanently mount slides with a coverslip.

  • Tested applications

    Suitable for: IHC-Fr, IHC-Pmore details

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 24 ml 72 ml 240 ml
    Antibody Blocker (40x) 2 x 15ml 1 x 50ml 1 x 100ml
    Aqueous Mounting Medium 1 x 15ml 2 x 18ml 1 x 120ml
    Blocker A 1 x 12ml 2 x 18ml 1 x 120ml
    Blocker B 1 x 12ml 2 x 18ml 1 x 120ml
    DAB Chromogen (20x) 1 x 1.5ml 1 x 2ml 1 x 6ml
    DAB Substrate 1 x 15ml 2 x 18ml 1 x 120ml
    DAB-Ni Substrate (20x) 1 x 1ml 1 x 2ml 1 x 6ml
    Hydrogen Peroxide (20x) 1 x 1ml 1 x 2ml 1 x 6ml
    Mouse HRP Polymer 1 x 6ml 1 x 18ml 1 x 60ml
    Mouse Primer 1 x 12ml 1 x 36ml 1 x 120ml
    Nickel Solution (7x) 1 x 3ml 1 x 6ml 1 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
    Rabbit HRP Polymer 1 x 12ml 2 x 18ml 1 x 120ml
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab183296 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.

Protocols

References

ab183296 has not yet been referenced specifically in any publications.

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