Good Antibody to Detect p-Foxo1/3 Activity

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:

Phospho-FoxO1 (Thr24)/FoxO3a (Thr32) Antibody

Catalog Number:

9464

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 mTORC2 activity in WT T cells upon T cell activation. Foxo proteins are a downstream node of mTORC2; therefore, we measure phosphorylation of these proteins to assess mTORC2 signaling.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Activated murine CD8+ T cells

Primary Incubation

1:2500 in 4%BSA/TBST 4 deg overnight with shaking

Blocking Agent

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

Secondary Incubation

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

Tertiary Incubation

None

Detection

UVP Imaging System

Results Summary

We are able to detect p-Foxo1/3 activity in our T cell lysates. The signal is quite strong when cells were stimulated with TCR activation.

DOI or PMID #

N/A

Additional Notes

Probe with total Foxo1 antibody to ensure correct band.

Image Gallery

Summary

The Good

Quite robust of a signal and can use diluted antibody multiple times.

The Bad

None

The Bottom Line

One of the few antibodies available to truly assess p-Foxo1/3 activity. Very widely used by scientists.

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