ß-Actin antibody for Western Blotting

Biochemistry
Stanford University
Senior Research Scientist

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Company:

Sigma-Aldrich

Product Name:

Monoclonal Anti-ß-Actin antibody produced in mouse

Catalog Number:

A5316-100UL

Browse Similar Products: Beta-Actin Western Blot Antibodies

Protein Kinase D is family of Ser/Thr kinases that regulate multiple biological functions including cell migration, proliferation, apoptosis, transcriptional activation, angiogenesis, and Golgi organization and vesicle trafficking. In the present study, the molecular basis of Protein Kinase D recruitment to the Golgi complex was investigated. I had discovered that class I and Class II ARF proteins and more specifically ARF1 can function as a tethering factor for Protein Kinase D at the Golgi complex. These conclusions were based on protein-protein interaction assays, colocalization experiments, and Golgi functional assays after knockdown down of Protein Kinase D in cells. Beta-actin antibody was used asa loading control for Western Blotting experiments. This antibody was used because many papers have cited this product.

Experimental Design and Results Summary

Applications

Western Blot

Sample

HeLa cell whole cell lysate

Primary Incubation

1:5000 dilution in blocking buffer; 1 hr at room temp

Blocking Agent

5% Milk powder in 1xTBS-Tween (0.1%)

Secondary Incubation

Goat anti-mouse IgG-HRP conjugated; 1:10000 dilution in blocking buffer; 1 hr at room temp;

Tertiary Incubation

None

Detection

ECL regular

Results Summary

Beta-actin antibody was used as a loading control to make sure that sample loading was uniform across lanes in a SDS-PAGE gel followed by Western Blotting. This antibody is very specific and probably incubation for 20-30 min would also be fine.

Additional Notes

None

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Summary

The Good

Specific signal and no background issues

The Bad

None

The Bottom Line

Perfect antibody for loading control determination in Western Blotting experiments

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