Great Human Lyn Antibody

December 11, 2015

Research
University of North Carolina at Chapel Hill
Laboratory Technician

Overall

Quality of Results

Ease-of-Optimization

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Company:

Cell Signaling Technology

Product Name:

Lyn (C13F9) Rabbit mAb

Catalog Number:

2796

Our lab investigates the role of kinases in several cancers, and we perform a handful of Western Blots each week to elucidate kinase activity in various cell lines. For the purposes of our research, we needed to detect endogenous levels of the human Lyn kinase in 786-0, RPTEC, and RCC4 cell lysates. We chose this CST antibody because of their reputable customer service and well-developed antibodies. Furthermore, the literature states that this antibody does not cross-react with any other Src-family members, which allows us to specifically measure this member of the Src kinase family.

Experimental Design and Results Summary

Applications

Western Blot

Sample

786-0, RPTEC, and RCC4 Cell Lysates

Primary Incubation

1:1000 Dilution Overnight in 5% BSA in 1X TBS-T at 4C

Blocking Agent

5% Milk in 1X TBS-T

Secondary Incubation

1:5000 dilution of HRP-conjugated Rabbit secondary in 5% Milk in TBS-T for 1 hour at RT

Tertiary Incubation

NA

Detection

ECL --> Autoradiography Film via HRP-Conjugate Secondary Antibody

Results Summary

We were able to accurately detect total Lyn kinase levels in human 786-0 cells and several other cancer cell lines with this antibody. Using autoradiography film, there was very little background and a strong signal with just a low-strength ECL mixture. By blotting with this antibody, we were able to accurately determine an shRNA-directed Lyn knockdown. Our lab prefers Cell Signaling Technologies antibodies for the reasons above, and we will continue to do so.

Additional Notes

Start out with a low-strength ECL mixture and move up as needed. The signal is medium-strength.

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Summary

The Good

Specific for Lyn, low background, moderate signal

The Bad

Expensive

The Bottom Line

If your research investigates Lyn protein levels, this antibody is a reliable method by which to detect them in human cell lysates. The price isn't cheap, but the extensive antibody validation and reputable customer service offered by Cell Signaling Technologies are worth it.

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