TripleStain IHC Kit: M&M&R on Human tissue (DAB, AP/Red & HRP/Green) (ab183286)

Overview

  • Product name

    TripleStain IHC Kit: M&M&R on Human tissue (DAB, AP/Red & HRP/Green)
  • Assay type

    Polymer Multiplex
  • Product overview

    Abcam's TripleStain IHC kit (ab183286) is designed to use with user supplied two mouse primary antibodies and one rabbit primary antibody to detect three distinct antigens on a single human tissue or cell samples. This kit has been tested on paraffin embedded tissue only; however it may also be used on frozen or freshly prepared monolayer cell smears.


    Triple staining uses traditional and non-traditional methods in immunohistostaining to reveal three distinct antigens and their co-expression on a single tissue. This kit supplies polymer enzyme conjugates: Rabbit HRP Polymer, Mouse HRP Polymer and Mouse AP Polymer with three chromogens, DAB Chromogen (brown); Permanent Red Chromogen (red); and Emerald Chromogen (green). Abcam's TripleStain IHC kit (ab183286) is a non-biotin system, avoiding non-specific binding caused by endogenous biotin. This kit has been optimized to have no cross detection when detecting two primary antibodies from the same host species using our 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 the user 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 Identifier 24 ml 72 ml 240 ml
    Antibody Blocker (40x) 2 x 15ml 1 x 50ml 1 x 100ml
    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
    Emerald Chromogen 1 x 15ml 2 x 18ml 1 x 120ml
    Mouse AP Polymer 1 x 6ml 1 x 18ml 1 x 60ml
    Mouse HRP Polymer 1 x 12ml 2 x 18ml 1 x 120ml
    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
    Plastic pipette 1 unit 1 unit 1 unit
    Rabbit HRP Polymer 1 x 6ml 1 x 18ml 1 x 60ml
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab183286 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.

Images

  • Immunohistochemical analysis of paraffin embedded human colon cancer tissue. Mouse anti-smooth muscle actin (red), rabbit anti-CEA (brown), mouse anti-PCNA (emerald).

Protocols

References

This product has been referenced in:

  • Cui G  et al. Up-regulation of FGF15/19 signaling promotes hepatocellular carcinoma in the background of fatty liver. J Exp Clin Cancer Res 37:136 (2018). Read more (PubMed: 29973237) »
See 1 Publication for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

TripleStain IHC KIT: M & M & R

Excellent Excellent 5/5 (Ease of Use)
Abreviews
A nice kit. It is easy to use and the instructions are easy to handle and to follow.
These are beautiful clear color results that you can also optically separate from each other. There was also no nonspecific staining due to double mouse staining. We used the kit on paraffin samples and performed a hematoxylin counterstain.

Abcam user community

Verified customer

Submitted Sep 02 2019

Question
Answer

Yes you should be able to use this this kit on an autostainer except for the emerald chromogen step.

Things to consider when you are setting up the instrument:



Please note that antibody concentration/titer may not be the same for manual and autostainer protocols.
You may need to optimize the time of primary antibody incubation. Especially if they are doing a mouse and rabbit combination.
To increase the staining I would add 10 to 15 minutes to the incubation time of the polymer to adjust for some of the autostainer will dilute the product as it travels through the tubing.
Remember that you will have to add a pause step to mix the Permanent Red AP chromogen since this product is mix just prior to application for optimal staining.
If you want to do double AP I would mix a second batch after the first batch has been applied.
The emerald chromogen should be done manually since it is water sensitive. Please read the protocol for best application.

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