Great Human Hif2a Antibody

December 01, 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:

Hif-2a Rabbit Monoclonal Antibody

Catalog Number:

7096

This is arguably the best antibody for human Hif2a that I have found on the market. We have been using it for years in lab, and have no reason to change our minds. With this antibody, we are able to detect Hif2a protein levels in cell lysates following hypoxia incubations with minimal background levels on autoradiography film. We prefer CST antibodies because of the extensive validation process and the ease of customer service if any issues arise.

Experimental Design and Results Summary

Applications

Western Blot

Sample

A549 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

Hif2a protein levels were detected in A549 cell lysates with minimal background upon overexposure and clean bands were obtained with correct ECL/FEMTO exposure protocols. In addition to presence/absence, relative ratios of band strength were also detected. We were able to prove Hif2a knockdown in cell lysates of A549 cells treated with 6 hours of 100 uM DFO using this antibody.

Additional Notes

None

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Summary

The Good

Clean, reliable, with great customer service from CST

The Bad

Requires 1:1 FEMTO:ECL for adequate exposure of bands

The Bottom Line

Buying an antibody from CST that has undergone extensive validation procedures was certainly worth it. I will continue to use this antibody for publication-quality western blots proving Hif2a knockdown or presence.

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