Western Blot of HeLa Whole-Cell Lysates Using Actin-Specific Antibody

February 20, 2019

Laboratory of t-RNA Transcription
Centre for Cellular and Molecular Biology
Research Associate

Overall

Quality of Results

Ease-of-Optimization

What do these ratings mean?
Write a Review

Company:

Cell Signaling Technology

Product Name:

β-Actin (13E5) Rabbit monoclonal antibody

Catalog Number:

4970

Our aim was to examine the protein levels of our work under untreated and bleomycin (which induces double-stranded breaks) treated HeLa cells. We extracted whole-cell lysates from cells using the 1xNETN buffer and loaded an equal amount of protein in both untreated and treated wells. Beta-actin was used as a loading control.

Experimental Design and Results Summary

Applications

Western Blot

Sample

HeLa whole cell lysate

Primary Incubation

1:5000, 1 hour at room temperature

Blocking Agent

5% skimmed milk, 1 hr at room temperature

Secondary Incubation

1:10000, anti-rabbit HRP conjugated antibody, 1 hr at room temperature

Tertiary Incubation

N/A

Detection

ECL reagent, Chemi doc

Results Summary

We observed a single specific band corresponding to actin at 45 kDa with western blot, and the levels were unchanged in both untreated and treated conditions in whole-cell lysates.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Excellent antibody for performing western blots

The Bad

N/A

The Bottom Line

Excellent antibody to perform western blots and east to optimize.

Share your experience with other scientists. Write a Review! »

Join the discussion