Western Blot of HeLa Whole Cell Lysates Using TSC2-Specific Antibody

October 08, 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:

Tuberin/TSC2 Antibody

Catalog Number:

3612S

The aim of our research was to study the role of Tuberin (TSC2) protein in regulating mTOR signaling in mammalian cells. We used TSC2 antibody to analyze the levels of TSC2 protein in HeLa cell lysates transfected with scramble and TSC2 siRNA by western blot.

Experimental Design and Results Summary

Applications

Western Blot

Sample

HeLa whole cell lysate

Primary Incubation

1:2000, overnight at 4°c

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 decreased levels of TSC2 levels in TSC2 siRNA-transfected cells compared to scramble siRNA-transfected cells. A single specific band at approximately 200KD was observed by western blot.

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

Specificity and easy to optimise

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