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Rabbit specific HRP/AEC (ABC) Detection IHC Kit (ab64260)

Submit a review Q&A (3)References (1)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti Rabbit HRP/AEC Detection Kit (ab64260)

    Overview

    • Product name

      Rabbit specific HRP/AEC (ABC) Detection IHC Kit
      See all HRP/AEC Detection Kit kits

    • Product overview

      Rabbit specific HRP/AEC (ABC) IHC Detection Kit ab64260 is a complete kit to detect a primary antibody raised in rabbit, using HRP-labeled-streptavidin and a biotinylated anti-rabbit secondary antibody.


      It is supplied with a full protocol and with peroxidase and protein blocking reagents, biotinylated secondary antibody, streptavidin-HRP, and AEC substrate.


      In the method:
      - a sample is incubated with an unconjugated primary antibody specific to the target antigen
      - a biotinylated secondary antibody then binds to the primary antibody
      - HRP-labeled streptavidin then binds to the secondary antibody
      - the HRP produces a colored substance at the site of primary antibody binding by reacting with AEC.

    • Notes

      For alternative AEC streptavidin-biotin IHC detection kits see:
      - anti-Mouse / anti-Rabbit HRP-AEC IHC kit ab93705
      - Anti-Mouse HRP-AEC IHC Kit ab64258

      For high sensitivity AEC HRP micro-polymer IHC detection kits see:
      - Anti-Mouse / anti-Rabbit HRP-AEC Micro-polymer IHC Kit ab236467
      - Anti-Rabbit HRP-AEC Micro-polymer IHC Kit ab236468

      Alternatively, see HRP-DAB IHC detection kits.

      Find more kits and reagents for antigen retrieval, blocking, signal amplification, visualization, counterstaining, and mounting in the IHC kits and reagents guide.

    • Tested applications

      Suitable for: IHC-Pmore details

    Properties

    • Storage instructions

      Store at +4°C. Please refer to protocols.
    • Components 15 ml
      AEC Single Solution 1 x 15ml
      Biotinylated goat anti-rabbit IgG(H+L) 1 x 15ml
      Hydrogen Peroxide Block 1 x 15ml
      Protein block 1 x 15ml
      Streptavidin Peroxidase 1 x 15ml

    • Research areas

    • Relevance

      An HRP/AEC Detection Kit is a labeled immunoenzymatic antigen detection system. Horseradish peroxidase (HRP) readily combines with hydrogen peroxide (H2O2) and the resultant complex can oxidize a wide variety of chromogenic substrates. AEC is 3-amino-9-ethyl-carbazole, a chromogenic substrate.

    • Alternative names

      • carbazole
      • Horseradish Peroxidase

    Applications

    The Abpromise guarantee

    Our Abpromise guarantee covers the use of ab64260 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-P
    Use at an assay dependent dilution.
    Notes
    IHC-P
    Use at an assay dependent dilution.

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti Rabbit HRP/AEC Detection Kit (ab64260)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti Rabbit HRP/AEC Detection Kit (ab64260)
      Human tonsil stained with anti-CD3 antibody using Anti Rabbit HRP/AEC Detection Kit (ab64260)

    References (1)

    Publishing research using ab64260? Please let us know so that we can cite the reference in this datasheet.

    ab64260 has been referenced in 1 publication.

    • Friedman O  et al. Immunological and inflammatory mapping of vascularized composite allograft rejection processes in a rat model. PLoS One 12:e0181507 (2017). IHC-P ; Rat . PubMed: 28746417

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-3 of 3 Abreviews or Q&A

    Question

    xxxx;

    As I told you on the phone, I am planning on performing IHC on fresh PBMC samples from sheep.

    Cells will be isolated from whole blood using a density gradient and will be placed on glass slides using a cytocentrifuge. At this point, we CAN fix them if needed, but we don’t have to. What do you recommend?

    Our primary antibody is affinity purified rabbit polyclonal. We care considering using your EXPOSE Rabbit specific HRP-AES detection kit (ab94361) – do you think this is the best kit to use, or would you recommend a different one for our purposes? We are attempting to stain a transmembrane protein.

    We are planning to counterstain using hematoxylin: do you think we would get the best contrast with the AEC kit or would you recommend using DAB?

    Also, does abcam recommend purchasing hematoxylin from any particular vendor?

    If ab64230 the appropriate mounting medium to use with this kit?

    Thanks so much for your help. I look forward to your response!

    Read More
    Answer

    Thank you for replying with that information.

    As I mentioned over the phone, I have spoken to the source of these kits and we do not have any experimental data using either the ACE or DABdetection kits (ab64260, ab64261, ab80437 and ab94361)for IHC on fresh PBMC samples, asallof these kits have only been tested on formalin fixed paraffin embedded tissue samples.Wewould expect that as long as the primary antibody binds the target as expected that both kits could be suitable foryour samples but as we have not tested it ourselves we would not be able to guarentee this.



    There should be good contrast if using the AEC or DAB kit with hematoxylin as counterstain. We counterstain with hematoxylin in-house regardless ofwhether DAB or AEC is used. The sensitivity betweenAEC and DABis similar. I would recommend using ab94361 or ab80437 if you expect the PBMC samples to contain any endogenous biotin.I am sorry but I cannot recommend a particular supplier of hematoxylin.

    And to answer your final question, the mounting media ab64230 should be suitable for your purposes.

    I hope this information has been of help. If you require any further information please do not hesitate to contact us again.

    Read More

    Question

    One of our customers wants to use ab93705 kit for IHC on Mouse samples.
    The secondary antibody included with the kit is anti Mouse/Rabbit, I'm concerned it might react with the mouse tissue and result in unspecific staining.
    Is the suitable for Mouse derived samples and would the included blocking reagent prevent unspecific staining? Is there a more suitable kit for this purpose?

    Read More
    Answer



    Our ab93705 (Mouse and Rabbit Specific HRP/AEC (ABC) Detection IHC Kit) is not suitable for use on mouse samples. The secondary antibody is a cocktail of biotinylated anti-mouse and anti-rabbit antibodies and the anti-mouse antibodies will react with endogenous Ig in the mouse samples. The blocking reagent will not prevent that staining.

    If the customer is using a rabbit primary antibody I would recommend that they use our ab64260 (Rabbit specific HRP/AEC (ABC) Detection IHC Kit) instead:

    https://www.abcam.com/Rabbit-specific-HRPAEC-ABC-Detection-IHC-Kit-ab64260.html (or use the following: https://www.abcam.com/Rabbit-specific-HRPAEC-ABC-Detection-IHC-Kit-ab64260.html).

    Read More

    Question

    What is the protein block?

    Read More
    Answer

    Thank you for contacting us. I can confirm the protein block has the following composition: PBS solution, pH 7.6 0.5% BSA 0.5% casein Less than 0.1% sodium azide I hope this information helps. Please do not hesitate to contact me should you have further questions.

    Read More

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