Good Phospho-STAT3 Antibody for Protein Quantification

Department of Pharmacology
Yale University
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

Cell Signaling

Product Name:

Phospho-Y705 STAT3 Antibody

Catalog Number:

9145S

Clone Number:

D3A7

Info:Supplier Page View Product Specs

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We knocked out an upstream JAK protein in the Cal27 cell line and wanted to examine downstream STAT3 signaling. We blotted JAK KO (KO) and control cell lines for Phospho-Y705 STAT3 in starvation and nutrient-rich cell culture conditions.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Cal27

Primary Incubation

1:1000 dilution of primary antibody in 1% BSA incubated overnight at 4 degrees celsius on a slow rocker.

Blocking Agent

5% milk

Secondary Incubation

1:1000 goat-anti-rabbit HRP conjugated antibody suspended in 1% milk for one hour at room temperature on a slow rocker.

Tertiary Incubation

N/A

Detection

Film

Results Summary

We found that the JAK KO cells had a decrease in Phospho-Y705 STAT3 signalling both in starvation and nutrient rich conditions.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Robust bands in expected size range.

The Bad

There is some limited background signal in the expected size range.

The Bottom Line

This antibody clearly detects Phos-Y705 STAT3 in the expected size range with limited background.

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