Rabbit specific HRP/DAB (ABC) Detection IHC Kit (ab64261)


  • Product nameRabbit specific HRP/DAB (ABC) Detection IHC Kit
    See all HRP/DAB detection kit kits
  • Product overview

    The reagents in this kit constitute a labeled streptavidin-biotin immunoenzymatic antigen detection system. This technique involves the sequential incubation of the specimen with an unconjugated primary antibody specific to the target antigen, a biotinylated secondary antibody which reacts with the primary antibody, enzyme-labeled streptavidin, and substrate-chromogen.

  • Notes

    DAB is a suspected carcinogen. Handle with care.

    Contains hydrogen peroxide

  • Tested applicationsSuitable for: IHC-P, IHC-Frmore details


  • Storage instructionsStore at +4°C. Please refer to protocols.
  • Components 15 ml
    50x DAB Chromogen 1 x 0.5ml
    Biotinylated goat anti-rabbit IgG(H+L) 1 x 15ml
    DAB substrate 1 x 15ml
    Hydrogen Peroxide Block 1 x 15ml
    Protein block 1 x 15ml
    Streptavidin Peroxidase 1 x 15ml
  • Research areas
  • RelevanceHRP/DAB detection kit is a immunoenzymatic antigen detection system for immunohistochemistry techniques.


Our Abpromise guarantee covers the use of ab64261 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 concentration.
IHC-Fr Use at an assay dependent concentration.

Rabbit specific HRP/DAB (ABC) Detection IHC Kit images

  • Immunohistochemical analysis of KLF4 and pVHL expression in colon cancer tissues. Sections from colon cancer and adjacent tissues were analyzed by immunostaining against KLF4 and pVHL. Staining without primary antibody served as negative control.

    Tissue sections were de-waxed with xylene and rehydrated through gradient ethanol into water. For antigen retrieval, sections were heated in citrate buffer (pH6.0) for 10 min at 95°C in a microwave oven. After cooling to room temperature, the sections were then digested with 0.05% trypsin for 10 min at 37°C. Endogenous peroxidase activity was quenched with 0.3% H2O2 in methanol for 30 min at room temperature. After PBS washes, nonspecific antibody binding was blocked by pre-incubating slides with 10% normal goat non-immune serum at 37° for 30 min. After blotting off the blocking serum, sections were incubated with primary antibody against LKF4 as well as primary antibody against VHL at 4° overnight. After PBS washes again, sections were incubated with biotinylated secondary antibody at 1:200 dilution for 30 min at room temperature. After incubating with Vectastain ABC reagent (Vector Laboratories, Inc., Burlingame, CA) for 30 min at room temperature, the sections were developed with Anti Rabbit HRP/DAB Detection Kit (ab64261). Sections were washed in running tap water and lightly counterstained with hematoxylin, followed by dehydration and coverslip mounting. Negative controls were obtained by omitting the primary antibody.

  • Human colon carcinoma fixed in 10% NBF for 24 hrs and stained with anti p53 antibody using Anti Rabbit HRP/DAB Detection Kit (ab64261).

References for Rabbit specific HRP/DAB (ABC) Detection IHC Kit (ab64261)

This product has been referenced in:
  • Lee JG & Wu R Erlotinib-cisplatin combination inhibits growth and angiogenesis through c-MYC and HIF-1a in EGFR-mutated lung cancer in vitro and in vivo. Neoplasia 17:190-200 (2015). IHC-P . Read more (PubMed: 25748238) »
  • Baiula M  et al. Mapracorat, a selective glucocorticoid receptor agonist, causes apoptosis of eosinophils infiltrating the conjunctiva in late-phase experimental ocular allergy. Drug Des Devel Ther 8:745-57 (2014). IHC-P . Read more (PubMed: 24959069) »

See all 12 Publications for this product

Product Wall

This issue may be result of sample preparation. It is oftentimes the case that the slides have not been inadequate deparaffinization which will lead to this type of staining. The link below outlines Abcam recommendations of deparaffinization: http:/...

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One drop of DAB chromogen is about 30 ul, and it is to be added to 1.5 ml of the substrate (50 drops). So the ratio is 1:50.

The mixture cannot be stored very long once you have made it. Ideally, you should mix it immediately before using it...

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I can confirm the Protein Block is a PBS solution with 0.5% Casein and 0.5% BSA.

I hope this information will be helpful for your customer.
Have a great day!

The lab sent me the following explanations:

The protocol for ab64261 reflects the sequence we follow in house. Personally I have done the endogenous peroxidase blocking step before antigen retrieval, after antigen retrieval and after the...

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Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has cau...

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I am sorry that I was unavailable when you called but am glad to hear from you. We would not suggest using 5% BSA instead of the protein block included with the kit. These blocking solutions are formulated to give the best result with the kit. I...

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It does look like endothelium is stained, as expected, but yes, the background is high. Did you include a negative control, staining with just the secondary detection kit, and not the primary? If that is completely negative, and shows no background, th...

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Thank you for contacting us. You will need an antigen retrieval step. Customers have reported success with a few different protocols but I recommend a heat mediated retrieval using a 10mM citrate buffer, pH 6.0. This should be applied to the sections b...

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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)f...

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Thank you so much for your call today and for letting us know about the trouble with the staining.

As we discussed, I'm sending a free of charge DAB avidin-biotin complex staining kit, ab64261, which should arrive on Monday on the order ***. ...

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