EZH1 Antibody Not Suitable for Fluorescence Immunocytochemistry

March 06, 2019

Cardiovascular Innovation Institute
University of Louisville
Research Coordinator

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Company:

MilliporeSigma

Product Name:

Anti-EZH1

Catalog Number:

abe281

In a freshly isolated rat microvessel prep from adipose tissue, anti-EZH1 antibody was localized mostly in the cytoplasm, not in the nucleus as expected. To be fair, this antibody has not been suggested for fluorescence immunocytochemistry by the manufacturer, and has not been verified for use in rat.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

Rat microvessels

Primary Incubation

1:1000 overnight

Blocking Agent

0.5% BSA and 10% NDS

Secondary Incubation

1:200 for 1 hour

Tertiary Incubation

N/A

Detection

Confocal microscopy

Results Summary

The antibody did show positive staining compared to isotype controls. However, EZH1 should be localized in the nucleus, and we found positive staining mostly in the cytoplasm.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

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Summary

The Good

Positive staining compared to isotype control

The Bad

Not localized where anticipated

The Bottom Line

While the staining protocol with this antibody "worked" in that there is positive staining compared to isotype controls, it was not primarily found in the nucleus, where EZH1 should be located.

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