Western blot review for anti-histone H3 (acetyl K27) antibody

Cardiology
Mount Sinai
Postdoc

Overall

Quality of Results

Ease-of-Optimization

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Company:

Abcam

Product Name:

Anti-histone H3 (acetyl K27) antibody

Catalog Number:

ab4729

This product was used to determine if changes in this histone modification are associated with endothelial to mesenchymal transition. This antibody was recommended by a coworker.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Chromatin isolated from untreated and TGFb2 treated HUVECs

Primary Incubation

1:3000 dilution in 5% skim milk in TBS-t overnight at 4 degrees

Blocking Agent

5% skim milk in TBS-t for 1 hour at room temperature

Secondary Incubation

1:10,000 dilution of anti-rabbit HRP conjugated secondary (thermo scientific Cat#31460) in 5% skim milk in TBS-t for 2 hours at room temperature

Tertiary Incubation

NA

Detection

Enhanced chemiluminescence

Results Summary

This histone modification was detected at 15kDa as a single band, and was decreased in HUVECs after treatment with TGFb2. Exposure time of 10 seconds with film.

Additional Notes

None

Related Categories

Image Gallery

Summary

The Good

Single band, very clean Western

The Bad

None

The Bottom Line

Would highly recommend this antibody.

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