• Product name
    DAB Substrate Kit
  • Product overview

    3,3'Diaminobenzidine (DAB) is a widely used chromogen for immunohistochemical staining. In the presence of peroxidase enzyme, DAB produces a brown precipitate that is insoluble in alcohol.This product is available in a two component form consisting of a liquid, refrigerator stable DAB Chromogen and DAB Substrate. Once the two components are combined, the reagent can be used for up to six hours, making it ideal for automated stainers.

  • Notes

    Contains DAB in buffer. Reagent is acidic and can cause burns if skin contact occurs.

    Contains Hydrogen Peroxide in DAB substrate buffer. Avoid contact with skin and eyes.

  • Tested applications
    Suitable for: IHC-Pmore details


  • Storage instructions
    Store at +4°C. Please refer to protocols.
  • Storage buffer
    Constituents: 100% Propylene glycol, 5% Hydrogen peroxide, DAB substrate buffer
  • Components 60 ml 125 ml 125 ml
    50x DAB Chromogen 1 x 2ml 1 x 4ml 1 x 4ml
    DAB substrate 1 x 60ml 1 x 125ml 1 x 125ml
  • Research areas
  • Relevance
    3,3' Diaminobenzidine (DAB) is a widely used chromogen for immunohistochemical staining. In the presence of peroxidase enzyme, DAB produces a brown precipitate that is insoluble in alcohol.
  • Alternative names
    • 3 3' Diaminobenzidine


Our Abpromise guarantee covers the use of ab64238 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.


  • Immuno-staining of formalin–fixed and paraffin-embedded liver sections with ab64238. Secontions were obtained from control (saline) and treated with CCl4+CT-N and CCl4+AD-N (left) and antibody control (right). 

  • Immunohistochemical analysis of healthy and colitis mouse colon sections (untreated and treated with enoxaparin) labeling claudin-4 with ab15104 at 1/200. ab7090, anti-rabbit immunoglobulin G conjugated to horseradish peroxidase (HRP) at 1/300 was used as the secondary antibody. ab64238 at 1/50 was used to develop the histological signal.

    Antigen retrieval was performed by incubating the sections for 10 minutes at 97°C in 1 mM EDTA buffer, pH 8.0 or 10 mM citrate buffer, pH 6.0.

    Control, C; untreated colitis, DSS; oral enoxaparin, OE; intraperitoneal injection of enoxaparin, IPE.

  • Human colon carcinoma stained with anti-p53 antibody using DAB Substrate Kit (ab64238)

  • Immunohistochemical analysis of mouse lung. Samples were fixed in 10% buffered formalin, paraffinized and sliced at 1.5 µm thick. Antigen retrieval was performed using ab64214 for the deparaffinized slices. Sections were blocked with 2% normal goat serum, PBS(-) and 0.1% Tween20. They were then incubated with the primary antibodies for 1 hour at 4°C and with secondary antibodies for 30 minutes at room temperature. The avidin-biotin-peroxidase complex method with peroxidase streptavidin and the DAB substrate kit ab64238 was performed.

    (A) A resected lung from a mouse sacrificed 28 days after SCL injection.

    (B) Haemotoxylin and Eosin staining showing the tumor composed of a central area with necrosis and a peripheral zone filled with SCLs.

    (C) Immunohistochemical staining for MMP-14 showing positive expression of MMP-14 in the peripheral zone of the tumor and negative in central zone.

    (D) Immunohistochemical staining weak staining of MMP-2 in the peripheral zone. 


This product has been referenced in:
  • Vasconcelos ACU  et al. A symptomatic swelling of the upper lip. Oral Surg Oral Med Oral Pathol Oral Radiol 125:107-111 (2018). Read more (PubMed: 29248422) »
  • Silva Dos Santos D  et al. Embryonic stem cell-derived cardiomyocytes for the treatment of doxorubicin-induced cardiomyopathy. Stem Cell Res Ther 9:30 (2018). Read more (PubMed: 29402309) »

See all 36 Publications for this product

Customer reviews and Q&As

This DAB substrate kit (ab94665) is easy to use, and the quality is good.

Liyong Zhang

Abcam user community

Verified customer

Submitted Aug 15 2017

Abreview for DAB substrate kit

Excellent Excellent 5/5 (Ease of Use)
This kit is streamlined to use at ease. The results is fantastic.

Liyong Zhang

Abcam user community

Verified customer

Submitted Aug 15 2017

IHC-P Mouse pancreas

Excellent Excellent 5/5 (Ease of Use)
Ki67 IHC staining on mouse pancreas, blocking with 10% BSA, 1 hr ab incubation RT

Abcam user community

Verified customer

Submitted Aug 16 2016

Thank you for contacting us.
The kit will work with the rabbit and mouse primary antibodies.
You can try citrate buffer pH for a start and Tris/EDTA in case the staining does not work. Please check the following for protocol and troubleshooti...

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Thank you for contacting us.
Heat mediated antigen retrieval method with microwave or pressure cooker is recommended for this ab.
The suitable detection system depends on the substrate you want to use. e.g. with DAB or AEC, we have following ...

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Thank you for contacting us again.

ab1002 Anti-Rabies Virus antibody looks very promising as it has been established for the use in IHC-P and should detect a wide range of rabies strains. As a starting dilution the customer can tr...

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Thank you for contacting us.

Here is the list with the requested items. However, please be aware that this is not a technical inquiry as all item could be found by using the search application on our website under the "reagents" ca...

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Thank you for contacting us. You are correct, these are the same product, just different sizes. The appropriate size will depend on how many slides you anticipate needing to stain. If you prepare the substrate/chromogen mixture as suggested in the prot...

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Thank you for your reply.

I looked over the protocol that you sent and it looks to be OK with me. I had a colleague look at it aswell and they thought it was good too.

If there is anything else I can help you with, or if you have an...

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Thank you for your reply and for providing those details.

I have now emailed you a proforma regarding the purchase of ab46667 and ab64238.

As for detailed step by step protocols, we do not have any that would meet your requirements,...

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1-10 of 21 Abreviews or Q&A


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