Preferred Actin-HRP Antibody

November 05, 2015

Research
University of North Carolina at Chapel Hill
Laboratory Technician

Overall

Quality of Results

Ease-of-Optimization

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Company:

Santa Cruz Biotechnology

Product Name:

HRP-Conjugated Actin Antibody (C-11)

Catalog Number:

sc-1615 HRP

Info: View Product View Product Specs

Browse Similar Products: Actin Antibodies

We perform a great deal of western blots to prove protein levels, and have been using this particular Actin-HRP antibody as the loading control for the majority of our western blots. With this antibody, we are able to accurately determine efficient well-loading and differences in protein concentrations between wells. Given how efficiently this antibody works, we plan to continue using this antibody for all western blots with actin used as the loading control.

Experimental Design and Results Summary

Applications

Western Blot

Sample

A549 cell lysates

Primary Incubation

1:5000 Dilution Overnight in 5% BSA in 1X TBS-T at 4C

Blocking Agent

5% Milk in 1X TBS-T

Secondary Incubation

None needed - already HRP-conjugated

Tertiary Incubation

NA

Detection

Autoradiography Film using ECL alone

Results Summary

This antibody yields consistently impressive results. Furthermore, the ease of use is great. I get reliably clean actin bands for each western blot, and the bands expose well with ECL alone. No need to worry about secondary antibodies - it's already HRP-conjugated. Even when left to dry, this antibody produces a strong and reliable chemiluminescent signal that is easily detected with autoradiography film.

Additional Notes

Be sure not to over-incubate. Less is more with this antibody.

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Summary

The Good

Quick and reliable. Already HRP-conjugated for ease of use.

The Bad

Different lots can bind differently

The Bottom Line

For the price and ease of use, this is a staple of our lab for assaying actin protein levels in cell lysates via western blot.

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