Great Antibody to Detect Total Foxo1 Protein Level for Immunoblotting

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:

FoxO1 (C29H4) Rabbit mAb

Catalog Number:

2880

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 based on Foxo signaling in WT T cells upon T cell activation. When we assess phospho-Foxo1(S256) activity, we also want to determine total Foxo1 to normalize activity to total protein.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Murine T cells

Primary Incubation

1:3000 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 easily able to detect total Foxo1 levels in our cell lysates. Even better since the pFoxo1 bands fall in the same area as the total Foxo1 bands.

DOI or PMID #

N/A

Additional Notes

None

Image Gallery

Summary

The Good

Very strong and reliable antibody. Can use diluted antibody multiple times if stored properly.

The Bad

None

The Bottom Line

Wonderful antibody to detect total Foxo1 protein levels with western blotting.

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