Measuring SIRT6 Knockdown in HeLa Cells by Western Blot

August 28, 2018

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

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Quality of Results

Ease-of-Optimization

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Company:

Abcam

Product Name:

Anti-SIRT6 antibody

Catalog Number:

ab88494

Our goal was to check the specificity of SIRT6 siRNA in knocking down SIRT6 levels in HeLa cells. We have transfected HeLa cells with a Scramble and SIRT6 siRNA for 48 hrs. Whole cell lysates were prepared after transfection and protein levels were analyzed by western blot using this SIRT6 antibody.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Control and SIRT6 siRNA treated HeLa whole cell lysates

Primary Incubation

1:5000, Overnight at 4'c

Blocking Agent

5% skimmed milk, 1 hr at room temperature

Secondary Incubation

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

Tertiary Incubation

N/A

Detection

ECL reagent, Chemi doc

Results Summary

A clear specific band was observed at approximately 40KD corresponding to SIRT6 in scramble siRNA treated HeLa cells and reduced significantly in SIRT6 siRNA treated HeLa whole cell lysates.A non-specific band appears approximately at 30KD which does not change in both siRNA treated cells.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

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Summary

The Good

Excellent antibody for performing western blots

The Bad

A non-specific band appears every time at 30KD

The Bottom Line

Overall an excellent antibody which specifically recognizes the SIRT6 protein in different cell lysates.

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