Beta Actin As A Western Blot Loading Control

January 09, 2017

Food Science and Human Nutrition
Colorado State University
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

Abcam

Product Name:

Anti-beta Actin

Catalog Number:

ab8227

Beta actin was used as a loading control for western samples to assure that protein loading is the same across the gel. This is especially important for proteins that do not get phosphorylated when they are activated and therefore cannot be tested against a total sample.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Total cell lysates of H4IIE rat hepatoma cells

Primary Incubation

1:1000 antibody in 1:3 Odyssey Block: TBST overnight, at 4 degrees C.

Blocking Agent

1:3 Odyssey Block: TBS for one hour at room temperature

Secondary Incubation

1:15000 IRDye800CW in 1:3 Odyssey Block: TBST

Tertiary Incubation

N/A

Detection

Licor Odyssey Clx

Results Summary

Beta actin is a simple and effective loading control. No optimization was necessary as it worked the first time and displayed a strong signal at 40kDa that was equal through all samples.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

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Summary

The Good

No background. Easy Optimization. Strong signal.

The Bad

Abcam is pricey but have a money back guarantee if the antibody does not work

The Bottom Line

This antibody is a great loading control with no variability throughout samples. Would recommend and will use again.

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