Phospho-p70 S6 Kinase (Thr389) (108D2) Rabbit mAb Antibody (for Western Blot) from Cell Signaling Techonology

August 08, 2014

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Women & Infants Hospital
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Company:

Cell Signaling Techonology

Product Name:

Phospho-p70 S6 Kinase (Thr389) (108D2) Rabbit mAb

Catalog Number:

#9234

This antibody was used in a study that investigated the role of Phospho-p70 S6 Kinase (Thr389) in WT and transfected cardiomyocytes. Several Phospho-p70S6K antibodies were tested in this study for various applications; the antibody reviewed here (Cat# 9234) gave the best results in Western Blot of Phospho-p70 S6Kinase (Thr389) on the expression of WT and stably transfected with Mutation cardiomyocytes.

Experimental Design and Results Summary

Applications

Western Blot

Sample

H9C2 rat cardiomyocytes stably transfected with WT and mutant plasmid

Primary Incubation

Phospho-p70S6Kinase(Thr 389) antibody (Cell Signaling, cat.no. 9234), incubated 2h at 4°C in Blotting-Grade Blocker (1:1000 dilution)

Blocking Agent

Blotting-Grade Blocker (BIO RAD, cat.no 170-6404)

Secondary Incubation

Anti-rabbit IgG, HRP-linked antibody (Cell Signaling, cat. no. 7074), incubated 2h incubation at room temperatureC (1:5000 dilution)

Tertiary Incubation

N/A

Detection

Amersham TM ECL Prime Western Blotting Detection reagent (GE Heathcare, cat. no. RPN2232), Image J software

Results Summary

This antibody worked excellently in Western Blt analysis of cardiomyocytes expression of Phospho-p70S6Kinase (Thr389). Only minimal background blotting control could be detected.

Additional Notes

This antibody could not be successfully used for other applications such as histochemistry or immunofuorescence staining experiments

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Summary

The Good

Highly specific in p-P70S6K (Thr389) analysis, low background

The Bad

Only useful for Western Blot

The Bottom Line

This is a very good antibody to detect p-P70S6K(Thr389) on cardiomyocytes cell lines. The minimal background blotting is tolerable when comparing to expression cells. However, this antibody might not be suitable to be used in immuno-fluorescence staining experiment.

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