Immunoblotting of ß-Actin for Loading Control

Dpt of Cell Bio
Havard Medical School
Postdoctoral Researcher

Overall

Quality of Results

Ease-of-Optimization

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Company:

Santa Cruz Biotechnology

Product Name:

β-Actin Antibody (AC-15)

Catalog Number:

sc-69879

Clone Number:

AC-15

This antibody was used as loading control for various experiment and used across various cell lines.

Experimental Design and Results Summary

Applications

Western Blot

Sample

NGN2-induced Neuron cells extracts

Primary Incubation

1:4000 1h incubation at room temperature, in 5% BSA diluted TBS+Tween (0.25%)

Blocking Agent

10% non-fat milk in TBS+Tween (0.25%) for 10 min

Secondary Incubation

1:20000, 1 h incubation at room temperature in 5% non-fat milk diluted TBS+Tween (0.25%)

Tertiary Incubation

Washed three times with TBS+Tween (0.25%) for 3 min each.

Detection

HRP-based chemiluminescent immunoblotting and x-ray film

Results Summary

This antibody detected a single band as expected and is well suited for loading control use across different samples. Works well in various cell line tested (HeLa, HCT116, Ramos, HEK293, U2OS, Neurons).

DOI or PMID #

10.1016/j.molcel.2018.03.012

Additional Notes

N/A

Related Categories

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Summary

The Good

Great sensitivity and specificity

The Bad

None

The Bottom Line

Awesome antibody. Highly recommended. Does the jobs in short incubation and is very specific.

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