Great Loading for Ribosomal S6 Protein for Immunoblotting

Oncology
Johns Hopkins School of Medicine
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

Cell Signaling Technology (CST)

Product Name:

S6 Ribosomal Protein (5G10) Rabbit mAb

Catalog Number:

2217

Clone Number:

Our lab studies the role of mTOR signaling in T cell such as mTORC1 and mTORC2 signaling during T cell differentiation and function. With immunoblot analysis, we assess mTOR activity in WT T cells upon T cell activation. We use this antibody to normalize total ribosomalS6 protein compared to pS6 (S235.236) activity in certain experiments.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Murine CD8+ T cells

Primary Incubation

1:10000 in 4% BSA/TBST 4 deg overnight with shaking

Blocking Agent

5% dry milk/TBST

Secondary Incubation

1:5000 anti-Rabbit HRP (CST) in 5% Milk/TBST 1hr Room Temp with shaking

Tertiary Incubation

None

Detection

ECL and UVP Imaging System

Results Summary

We are able to detect total ribosomal S6 levels in our lysates. The signal is quite strong depending on experimental conditions.

DOI or PMID #

N/A

Additional Notes

Dilute appropriately

Image Gallery

Summary

The Good

Reliable and robust antibody for ribosomal S6.

The Bad

None.

The Bottom Line

Great antibody to detect total ribosomal S6 protein when conducting western blots for mTOR signaling.

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