Robust Antibody to Detect mTORC1 Activity through pS6 Signaling

Oncology
Johns Hopkins School of Medicine
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

Cell Signaling Technology

Product Name:

Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP Rabbit

Catalog Number:

4858

Our lab studies the role of mTOR signaling in T cell such as mTORC1 signaling during T cell differentiation and function. With immunoblot analysis, we routinely assess pS6 (S235.236) activity as a direct readout of mTORC1 signaling in WT T cells upon T cell activation.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Murine CD8+ T cells

Primary Incubation

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

Blocking Agent

5%milk/TBST for 1hr room temperature with shaking

Secondary Incubation

1:5000 anti-Rabbit HRP (CST) in 5%milk/TBST for 1hr room temperature with shaking

Tertiary Incubation

None

Detection

UVP Imaging System

Results Summary

We are able to detect mTORC1 activity by assessing pS6 (S235.236) in WT T cells upon stimulation through immunoblot analysis.

DOI or PMID #

N/A

Additional Notes

Titrate antibody since it can be quite bright depending on cell type and activity

Image Gallery

Summary

The Good

Great monoclonal antibody to detect mTORC1 signaling. Can reuse antibody at least 10 times plus.

The Bad

None

The Bottom Line

Easy-to-use antibody to analyze mTORC1 activity via western blotting.

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