Reporter System For Immunohistochemistry (IHC) With Rabbit Primary Antibodies (Abs)

March 09, 2017

Albert Einstein College of Medicine
Microbiology & Immunology
Post-doctoral Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

Vector Laboratories

Product Name:

ImmPRESS anti-Rabbit Ig Reagent Kit

Catalog Number:

MP-7401

Our lab performs IHC on mouse brain sections, and we stain for many different antigens of interest. The primary Abs that we use come from various species, including rabbits, rats, and goats. ImmPRESS's anti-rabbit Ig reagent kit provides sensitive, reliable recognition of our primary staining (rabbit Ab), ultimately yielding low-background signal and specific results.

Experimental Design and Results Summary

Application

IHC

Starting Material

Mouse brain tissue sections (embedded or frozen)

Protocol Overview

Following incubation of each section/slide with the (rabbit) primary Ab, perform wash steps with PBS. Add the ImmPRESS reagent (anti-rabbit Ig) to each section/slide. We typically incubate our sections for 30-45 minutes. Perform wash steps with PBS. Incubate each section/slide with a peroxidase substrate solution (we use DAB by Dako). Once the desired stain intensity develops, rinse with water, perform a counterstain, and mount each section/slide.

Tips

Optimize on controls, discarded sections prior to use on experimental samples. Dilute primary Ab in the serum included in the kit.

Results Summary

The results of each IHC experiment should be determined through control staining and comparison of experimental tissues to controls with known patterns or absence of staining.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Easy to use, quick/time efficient protocol, consistent results.

The Bad

Expensive, optimize on controls before using on experimental samples.

The Bottom Line

I highly recommend the anti-rabbit Ig ImmPRESS reagent kit for IHC as it is time efficient, easy to use, and provides consistent results. The micropolymer system provides us with both sensitive and specific detection of our antigens of interest in mouse brain sections (we typically use embedded sections, but the protocol indicates that frozen sections can also be used).

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