Best TYK2 Antibody

December 14, 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:

Tyk2 Antibody

Catalog Number:

#9312

Our lab aims to elucidate the role of kinases in several different types of human cancers, and we are often performing Western Blots to determine kinase protein and activity levels in various cell lines. Using new genome editing technology, we aimed to knockout the TYK2 kinase in human 786-0 cells, and used this antibody to validate our results. We generally look to Cell Signaling Technologies first for antibodies because of their extensive validation procedures and prompt customer service.

Experimental Design and Results Summary

Applications

Western Blot

Sample

786-0 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/FEMTO --> Autoradiography Film via HRP-Conjugate Rabbit Secondary Antibody

Results Summary

Using this antibody, we were able to provide evidence of TYK2 knockdown in 786-0 cell lysates. This allowed us to draw further conclusions about the influence of TYK2 upon renal cell carcinoma. Using autoradiography film and 1:1 mixture of ECL and FEMTO, there was very little background and a strong signal. A single non-specific band occurred above all samples, but it is well above ~134 kD and does not interfere with the principal band. Around 15-30 seconds was sufficient to produce bands on film that were strong but not overpowering.

Additional Notes

May be detected using just ECL for exposures of 2+ minutes

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Summary

The Good

Strong signal, low background, and a reliable manufacturer

The Bad

Expensive

The Bottom Line

In research, the value of time is often underestimated. While the price tag is expensive, not having to optimize the experiment, try multiple antibodies, or fumble around with bad customer service is well worth it. I highly recommend this particular antibody for total TYK2 kinase protein detection, and have used it for years without issue.

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