Not the Best p53 Antibody

Physiology
University of Tennessee HSC
Postdoctoral Researcher

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Company:

MyBioSource.com

Product Name:

p53 Antibody

Catalog Number:

MBS395752

Clone Number:

N/A

To test the time course effect of particular growth factor on p53 expression in mouse, the primary cell lines were growth-arrested for 12 hrs. in medium devoid of FBS. The synchronized cells treated without or with growth factor, prepared protein extracts, and an equal amount of proteins were resolved in 10%SDS-PAGE. The resolved proteins were then western transferred in a nitrocellulose membrane and probed with p53 specific antibody. Although, MyBioSource.com supply a polyclonal antibody, the selection of Ab was based on the merits indicated in antibody flyer.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Mouse primary cell whole cell lysate

Primary Incubation

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

Blocking Agent

skimmed milk (5%) prepared in 1x TBST

Secondary Incubation

Goat anti-sheep IgG HRP, dilution (1:4000) in 5% skimmed milk prepared in 1x TBST, 2hr at RT

Tertiary Incubation

None

Detection

ECL detection in X-Ray film

Results Summary

p53 from MyBioSource.com was a polyclonal antibody and it has detected a very faint band of p53 protein. Interestingly, a non-highly specific band with high Intensity was observed at higher molecular weight.

DOI or PMID #

None

Additional Notes

none

Image Gallery

Summary

The Good

Low specificity of Ab for p53

The Bad

Antibody highly cross-reacted with other nonspecific epitopes

The Bottom Line

Bad antibody for p53 detection

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