Total s6 Ribosomal Protein

Food Science and Human Nutrition
Colorado State University
Graduate Student

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Company:

Cell Signaling

Product Name:

S6 Ribosomal Protein

Catalog Number:

2217

The general aim of this project was to assess protein synthesis in a variety of treatment groups using ribosomal protein s6 (rps6), a downstream component of the MTORC1 pathway. To do this we compared phosphorylated rps6 to total rps6 using western blotting.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Total cell lysates of rat hepatoma cells

Primary Incubation

1:2000 antibody in 0.1% tween and 1:3 odyssey block: 1X TBS at 4 degrees C overnight.

Blocking Agent

1:3 odyssey block:1X TBS

Secondary Incubation

1:15,000 Li-Cor IRDye 800CW, 0.1% tween, 0.02% SDS, in 1:3 odyssey block: TBS at room temperature for 1 hour

Tertiary Incubation

N/A

Detection

Li-Cor Odyssey infrared Imaging system

Results Summary

There was a large distinction between samples that were phosphorylated (indicating active protein synthesis) and samples that were not phosphorylated (indicating halted protein synthesis). These results were consistent with another experiment performed to observe active protein synthesis via phosphorylation

Additional Notes

None

Related Categories

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Summary

The Good

Dilution from protocol worked well the first time and two times after. Have a lot of confidence with this antibody

The Bad

None

The Bottom Line

Had no issues in detecting total rps6 in this rat hepatoma cell line.

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