DAB+ Kit For Chromogen Development (Immunohistochemistry)

February 06, 2017

Albert Einstein College of Medicine
Clinical Research
Pre-doctoral researcher

Overall

Quality of Results

Ease-of-Optimization

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Company:

Invitrogen

Product Name:

DAB-Plus Substrate Kit

Catalog Number:

00-2020

We use Invitrogen's DAB-plus substrate kit to develop human brain sections stained for various immune cells. The protocol outlined by Invitrogen is simple and reliable, and once each protocol has been optimized we observe consistent, reliable results for each batch of sections that we stain. We selected the DAB+ kit because it includes an enhancer that produces a darker color than DAB alone, providing greater contrast. The inclusion of the enhancer in the kit is useful for some of our protocols, but we do not use it in every IHC staining protocol.

Experimental Design and Results Summary

Application

Chromogen development in IHC

Starting Material

Human tissue sections (following primary and secondary Ab applications)

Protocol Overview

Prepare the DAB reagents according to the manufacturer's guidelines.Following incubation with a secondary Ab-peroxidase conjugate, incubate each section in the DAB solution and monitor color development. Once desired color is achieved, rinse each section in distilled water and subsequently place the slides in the enhancer (if applicable to the protocol) for optimized amount of time. Rinse each section in distilled water, counterstain, and dehydrate/coverslip.

Tips

Optimize the protocol on control tissues prior to using prescious, experimental samples.

Results Summary

Successful staining can be determined by microscopy or a slide scanner. It is important to include control samples with known staining patterns to assure the utmost control of quality.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Relatively inexpensive, time efficient, simple protocol, reliable.

The Bad

Must thoroughly optimize prior to use.

The Bottom Line

I highly recommend this product to individuals who perform a lot of IHC on human sections. The kit is easy to use and the reagents provide consistent, reliable stain development.

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