Methyl Green for Immunohistochemistry

July 21, 2017

SUNY Downstate Medical Center
Department of Cell Biology
PhD Candidate (Graduate Student)

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Company:

Vector Laboratories

Product Name:

VECTOR Methyl Green

Catalog Number:

H-3402

We study the expression of a newly discovered receptor (TCblR) and another protein megalin in embryonic mouse brain using immunohistochemistry.

Experimental Design and Results Summary

Application

Immunohistochemistry

Starting Material

Paraffin embedded embryonic mouse brain sections

Protocol Overview

After immunostaining, we incubated the slides with sections on hot plate with temp. of 55-60 degree C. We covered the sections with methyl green and kept them for incubation for 30 minutes, adding more methyl green after every 10 minutes. After incubation, we rinsed the slides in tap water to get rid of excess methyl green. Then we dehydrated the slides in increasing concentration of ethanol, 70-100%, cleared in xyelenes and mounted using permount.

Tips

Dehydration after methyl green should be quick else the stain goes away

Results Summary

We got a nice blue-green color of nuclei in the brain which contrasts very well with DAB staining.

DOI or PMID #

28351841

Additional Notes

N/A

Image Gallery

Summary

The Good

Amazing color and contrast with DAB

The Bad

Goes bad pretty fast

The Bottom Line

The product works very well and gives amazing counter-staining and the pictures look amazing. However, it goes bad before it's expiry (storing at room temperature in dark) and doesn't work after that. For it's price, its expensive to keep buying it, when we get to use only less than a half container before it stops working.

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