Good Antibody for Interrogating mTOR Signaling (P-S6K1)

Oncology
Johns Hopkins University
Postdoctoral Researcher

Overall

Quality of Results

Ease-of-Optimization

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Company:

Cell Signaling Technology

Product Name:

Phospho-p70 S6 Kinase (Thr389) (D5U1O) Rabbit

Catalog Number:

97596

My research focus is on understanding the role of the immune system in regulating blood glucose and adipose tissue. For this experiment, mouse CD4+ T-cells were stimulated or stimulated with rapamycin. Rapamycin treatment inhibited S6K1 phosphorylation. P-S6K1 is a good readout for mTOR signaling.

Experimental Design and Results Summary

Applications

Western Blot

Sample

CD4+ mouse T-cells

Primary Incubation

4% BSA in TBST overnight at 4°C

Blocking Agent

5% milk in TBST

Secondary Incubation

5% milk in TBST for 2 hours at room temperature

Tertiary Incubation

N/A

Detection

ECL

Results Summary

Mouse CD4+ T-cells were non stimulated (left 4), stimulated (middle 4) or stimulated with rapamycin (right 4). Rapamycin treatment inhibited mTOR signaling by decreasing the phosphorylation of S6K1.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Nice clear band

The Bad

Weak signal

The Bottom Line

This is a good antibody to interrogate mTOR signaling. However, the signal can be a little weak.

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