Good Antibody To Detect Activated Src

May 01, 2018

The Cell and Molecular Biology
The Scripps Research Institute
Assistant Professor

Overall

Quality of Results

Ease-of-Optimization

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Company:

Cell Signaling Technology

Product Name:

Phospho-Src Family (Tyr416) Antibody

Catalog Number:

2101

The activity of the Src protein tyrosine kinase is important for regulation of cell growth, including proliferation of tumor cells. To become active, Src should be phosphorylated at Tyr416. Rabbit antibody #2101 detects phosphorylated tyrosine 416 in Src protein. The extent of Src activation is determined by comparing the levels of Tyr416-phosphorylated Src versus levels of total Src.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Lysates of human tumor cells; 20 microgram per lane

Primary Incubation

Antibody at a final dilution 1:1000, ON incubation with gentle rocking in a cold room

Blocking Agent

5% BSA in PBS-0.05% Tween 20

Secondary Incubation

Goat anti rabbit IgG conjugated with HRP, 1:3,000; 1 hr; RT; gentle rocking

Tertiary Incubation

N/A

Detection

West Pico

Results Summary

This rabbit anti Tyr416-Src antibody nicely detects a protein band with an apparent mol. wt. of ~61 kDa, expected for Scr protein.

DOI or PMID #

N/A

Additional Notes

The image demonstrates different levels of active (phosphorylated) Src in the lysates of human tumor cells containing similar levels of total Src.

Image Gallery

Summary

The Good

The rabbit anti Tyr416-Src appears to specifically detect activate Src in human tumor cells.

The Bad

None detected

The Bottom Line

Rabbit antibody #2101 from cell Signaling is a good and reliable antibody to detect activated Src in human tumor cells by western blotting.

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