Great Antibody to Detect PI3K Signaling Readout through pAKT

May 17, 2019

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-Akt (Thr308) (D25E6) XP® Rabbit mAb

Catalog Number:

13038

Clone Number:

Our lab studies the role of PI3k-mTOR signaling during T cell differentiation and function. We perform immunoblotting analysis of this signaling. For PI3K signaling, we assess pAKT (T308) activity mediated by PDK1 since this is the first readout in early signaling.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Activated Murine 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

With immunoblotting analysis, we are able to detect PI3K signaling based on pAKT (T308) activity upon T cell stimulation.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Robust and reliable antibody. Can reuse diluted antibody multiple times.

The Bad

Careful if using pAKT (S473) antibody since same size and patterns at times. Strip away well either one.

The Bottom Line

Great antibody to detect PI3K signaling with immunoblotting analysis.

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