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Chromogenic substrate for reporter enzyme


Item Cat No.: 00060

Application: Chromogenic substrate for IHC labeling

Reactivity: Tissue and Cell culture

100 ml size

Chromogenic substrate can be catalyzed by reporter enzyme used in IHC or in situ hybridization to form a colored precipitate as indicator of protein or RNA localization.

The choice of chromogen used is determined by the enzyme conjugated to an antibody as well as the color that chromogen generates. Because FFPE samples are post-stained by hematoxylin to label nuclei in purple to blue color, the chromogen’s color needs to have good contrast from hematoxylin, and is therefore in the range of red to brown color.

The following table summarizes the color variations of commonly used chromogens.

Enzyme Chromogen Precipitated color
Horseradish peroxidase (HRP) AEC Red
Horseradish peroxidase (HRP) DAB Brown
Horseradish peroxidase (HRP) TMB Blue
Alkaline phosphatase (AP) BCIP/NBT Blue
Alkaline phosphatase (AP) BCIP/TNBT Purple

Each substrate solution contains a mixture of substrate compounds and is ready to use. Apply substrate solution to tissue or cell samples until desired color is achieved (1-10 mins). Rinse off substrate with PBS or any other wash buffer.

Storage condition: ambient temperature

For Research Use Only. Not for use in clinical diagnostics.

Additional information

Reporter enzyme

Horseradish peroxidase, Alkaline phosphatase




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"I am really impressed with your approach. We tried multiple times previously to create monoclonal and polyclonal antibodies to claudin-2 and MLCK1. We have had limited success generating polyclonals and no success generating monoclonals. You have generated outstanding monoclonals to both. I look forward to continuing to work with you."

Jerrold R. Turner, M.D., Ph.D.

Brigham and Women’s Hospital | Harvard Medical School

"The polyclonal antibody you generated for KIAA0408 is stunning! KIAA0408 is a novel cilium molecule that has never been studied. So, clearly there will be a lot of demand for it as we have discovered a very interesting finding and the story will be published in a high impact journal. I am strongly inclined to generate monoclonal antibody for this protein too and we should think about patenting it."

Univ.-Prof. Jay Gopalakrishnan PhD

Heinrich-Heine-Universität | Universitätsklinikum Düsseldorf

"Your ARL13B antibody works beautifully!!! We’re so happy to have a cilia-specific antibody made in rat! I can send you high resolution images to be posted on your website."

Julie Craft Van De Weghe, PhD

School of Medicine | University of Washington

"The assay is a homophilic interaction mediated cell adhesion on purified protein (in this case, immobilized purified Pcdhga9 to Pcdhga9 expressed on cell surface). Compared to control, cell adhesion is reduced in the presence of Pcdhga9 monoclonal antibody supernatants!"

Divyesh Joshi, PhD

School of Medicine | Yale University

"The rabbit hybridoma supernatants of anti-APOBEC3 project are tested positive by ELISA, and we are very happy about it! We previously tried a company, Abclone. Their Project "A" has immune response that is <10,000 titer in antiserum, which would explain why there is no positive mAb after fusion. Their project "B" didn't have any immune response in rabbit."

Harshita B Gupta, PhD.

School of Medicine | UT Health San Antonio

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