Western Blot Using Rictor-Specific Antibody

November 20, 2018

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

Overall

Quality of Results

Ease-of-Optimization

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Company:

Cell Signaling Technology

Product Name:

Rictor (D16H9) Rabbit monoclonal antibody

Catalog Number:

9476S

Our aim was to examine the levels of histone acetylation in rictor-knockdown cells. We transfected HeLa cells with the scramble and rictor-specific siRNA and whole-cell lysates were successfully analyzed by western blot using rictor-specific antibody. Antibody is easy to optimize and gives a single specific band.

Experimental Design and Results Summary

Applications

Western Blot

Sample

HeLa whole-cell lysate

Primary Incubation

1:3000, Overnight at 4°C

Blocking Agent

5% skimmed milk, 1 hr at room temperature

Secondary Incubation

1:10,000, 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 rictor at a 200KD molecular weight. We observed decreased levels of rictor in cell lysates transfected with rictor-specific siRNA.

DOI or PMID #

29143563

Additional Notes

N/A

Image Gallery

Summary

The Good

Excellent antibody for performing western blots

The Bad

N/A

The Bottom Line

Easy to optimize and gives a single specific band in western blot

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