Great Antibody for Interrogating mTOR Signaling

March 12, 2019

Oncology
Johns Hopkins University
Postdoctoral Researcher

Overall

Quality of Results

Ease-of-Optimization

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Company:

Cell Signaling Technology

Product Name:

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

Catalog Number:

5364

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 and probed for P-S6 (middle 4). Rapamycin treatment inhibited the signaling (right 4).

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 in the presence of rapamycin (right 4). Activation increased P-S6 and mTOR signaling which was inhibited by rapamycin treatment.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Great signal detection

The Bad

Need to dilute antibody because very bright signal

The Bottom Line

Great antibody for interrogating P-S6 and mTOR signaling. You just have to use a high primary antibody dilution to prevent bleaching the membrane.

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