Mouse Monoclonal For Stat3

April 27, 2017

Biochemistry and Molecular Medicine
UC Davis
Graduate Student Researcher

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Quality of Results

Ease-of-Optimization

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Company:

Cell Signaling Technology

Product Name:

Stat3 Antibody

Catalog Number:

9139S

I was trying to detect the levels of Stat3 protein found in cells post-transfection of a drug in question. Stat3 is involved in the JAK-STAT signaling pathway, commonly indicative for cell proliferation and survival. I chose to purchase this antibody because of its specificity as a monoclonal.

Experimental Design and Results Summary

Applications

Western Blot

Sample

HEK293T Mammalian Cells

Primary Incubation

1:1000, 4C overnight

Blocking Agent

5% Milk

Secondary Incubation

Anti-mouse IgG, 1:3000 for 2 hours, room temperature

Tertiary Incubation

ECL Reagent, BioRad

Detection

ChemiDoc Imager, Hi-Sensitivity

Results Summary

Stat3 protein levels were downregulated by drug treatment, and confirmed experimentally in triplicates (first three lanes with treatment). The band was located near 100 kDa, near the reported molecular size from Cell Signaling of 79/86 kD.The monoclonal was specific, showing only one band at the expected size.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

This monoclonal is truly specific and generates beautiful bands.

The Bad

It's recommended to dilute the antibody in 5% milk, which means that re-use is limited.

The Bottom Line

I would definitely get a monoclonal antibody, as it is really hard to determine effect on the protein level with multiple bands. Smearing also makes quantification difficult. It is worth it to titer for the specific cell line you're using, but 1:1000 is definitely a good starting point.

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