Immunodetection of Cdk4 in Human Cell Line Using Abcam Antibody

Physiology
University of Tennessee HSC
Postdoctoral Researcher

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Company:

Abcam

Product Name:

Anti-Cdk4 antibody

Catalog Number:

ab226474

Clone Number:

Polyclonal

Browse Similar Products: Cdk4 Western Blot Antibodies

On-target siRNA transfected human cells treated with a growth factor for a specific time point were subject to screen the effect of siRNA on growth factor-induced post-translational modification of a protein kinase. After that, the same blot was stripped for IgG and normalise with cdk4 levels. For this after blocking the IgG removed membrane with 2% skimmed milk, the membrane was probed for cdk4 obtained from Abcam. Abcam antibody flyer showed good detection of target protein, so it was selected.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Human whole-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 Ab, dilution (1:4000) in 5% skimmed milk made in 1x TBST, 1hr at RT

Tertiary Incubation

None

Detection

Enhanced chemiluminescent detection in X-Ray films

Results Summary

Antibody from Abcam has detected bright and intense band at nearly 90 kDa, 60 kDa and 34 kDa. However, the lowest molecular weight protein band and 34 kDa was cdk4.

DOI or PMID #

N/A

Additional Notes

None

Related Categories

Image Gallery

Summary

The Good

Detected highly specific band of cdk4 at nearly 34 kDa

The Bad

Two non-specific bright bands at 90 kDa, 60 kDa were also observed

The Bottom Line

Good antibody for cdk4 detection

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