Average Antibody For The Immunodetection Of p300 In Human Cells

Physiology
University of Tennessee HSC
Postdoctoral Researcher

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Company:

Cell Signaling Technology

Product Name:

p300 (D1M7C) Rabbit mAb

Catalog Number:

70088

Clone Number:

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Mouse cells were treated with growth factor for a different period's cells were lysed in RIPA buffer, and proteins were extracted. The extracted proteins were resolved in 8% denaturing gel and western transferred in the nitrocellulose membrane. Further, the membrane was probed with p300 antibody to test the effect of growth factor treatment on p300 expression levels. The CST p300 antibody selection was based on our previous experience.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Mouse cell protein lysate

Primary Incubation

Dilution (1:1000) in 5% skimmed milk prepared in 1x TBST, overnight incubation at 4 degree Celsius

Blocking Agent

5% skimmed milk made in 1x TBST

Secondary Incubation

Goat anti-rabbit IgG HRP, Dilution (1:4000) in 5% skimmed milk made in TBST, 1hr at RT

Tertiary Incubation

None

Detection

Enhanced chemiluminescent detection in X-Ray film

Results Summary

A band of p300 protein was detected by this antibody. However, the non-specificity of this antibody is a concern.

DOI or PMID #

Additional Notes

None

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Summary

The Good

Although antibody detects p300 band, nonspecific bands were also detected.

The Bad

None

The Bottom Line

Average antibody for detecting p300 in mouse cells

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