Great Monoclonal Antibody to Detect all Akt Isoforms

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:

Akt (pan) (C67E7) Rabbit mAb

Catalog Number:

4691

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 mTOR activity in WT T cells upon T cell activation. When we assess phospho-Akt (S473) activity, we also want to determine total Akt to normalize activity to total protein.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Murine T cells

Primary Incubation

1:2000 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 total AKT levels in T cell lysates to normalize with pAKT S473 activity.

DOI or PMID #

N/A

Additional Notes

Good antibody to reuse 10 plus times if stored properly

Image Gallery

Summary

The Good

Good reliable and robust antibody.

The Bad

None

The Bottom Line

Great antibody to detect all Akt isoforms.

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