Actin for Normalizing Western Blot

Oncology
Johns Hopkins School of Medicine
Lab Technician

Overall

Quality of Results

Ease-of-Optimization

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Company:

Cell Signaling Technology

Product Name:

β-Actin Rabbit mAb

Catalog Number:

4970

Our lab is interested in comparing the mTOR, epigenetic, and metabolism signals of T cells in multiple media conditions with varying concentrations of glutamine. We use different concentrations to study the effects of the glutamine in the T-cell signaling. This antibody was used to normalize our bands. It is reliable and the signal is strong.

Experimental Design and Results Summary

Applications

Western Blot

Sample

CD8 T cells

Primary Incubation

1:5000 in 4°C overnight

Blocking Agent

5% nonfat dry milk

Secondary Incubation

Anti-rabbit IgG HRP-linked

Tertiary Incubation

N/A

Detection

ECL Pico

Results Summary

The actin is always very bright, sometimes too bright; therefore, we diluted down the recommended 1:1000 from CST to 1:5000 for our purpose. The bands are always clear and easy to read. We use the actin level to normalize across the wells of the band.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Very robust and bright antibody to normalize our bands

The Bad

None whatsoever

The Bottom Line

The actin never fails to show up whenever we expose our western blots. It is extremely reliable and strong, never nonspecific, and the bands are always consistent.

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