Expression of Phospho-S6 Ribosomal Protein decreased in acute Beta-adrenergic receptor antagonists treated mice

April 03, 2015

Pediatrics
Women & Infants Hospital
research Scientist

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Company:

Cell Signaling Technology

Product Name:

Phospho-S6 Ribosomal Protein (Ser240/244) (D68F8) XP® Rabbit mAb

Catalog Number:

5364

This antibody was used in a study that investigated the role of Phospho-S6 protein in acute beta-adrenergic receptor antagonist treated C57B6 mice. The aim of this project is to investigate the role of Beta-adrenergic receptors which are involved with cardiomyocytes proliferation. 2.5 days post pups were injected with Propranolol (10 mg/kg) or saline for 30 minutes in this study, then the hearts were collected and Western blot was performed; the antibody reviewed here (cat#: 5364, Cell Signaling) gave the best results in the Western blot study.

Experimental Design and Results Summary

Applications

Western Blot

Sample

The hearts from P2.5 post mice pups

Primary Incubation

Phospho-S6 Ribosomal Protein (Ser240/244) (D68F8) XP® Rabbit mAb (cell signalling, cat.no. #5364, incubated 24hs at 4°C in blocking reagent (1:1000 dilution)

Blocking Agent

5% blotting -Grade Blocker ( Bio-Rad, cat#: 170-6404) in TBST buffer

Secondary Incubation

Anti-rabbit IgG, HRP-linked antibody (Cell signalling, cat#:7074P2), incubated 2h incubation at room temperature (1:5000 dilution)

Tertiary Incubation

NA

Detection

Western blot

Results Summary

This antibody worked excellently in Western blot tests on mouse hearts. Almost no background was tested on the developed films.

Additional Notes

This antibody is easy and repeatable in Western blot, even after stripping the membrane 4 times, the signal can also be detected .

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Summary

The Good

Highly specific in Western blot analysis, low background

The Bad

A little expensive

The Bottom Line

This is a very good antibody to detect Phospho-S6 Ribosomal Protein expression in mouse hearts. The minimal background testing is tolerable when compared to expression of the proteins.

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