Great Human Hif1a Antibody

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-1a Antibody

Catalog Number:

3716

Info: View Product View Product Specs

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Accurate detection of human Hif1a protein in cell lysates is crucial to our research, and this antibody has continued to perform well for years of research in our lab. It is our go-to Human Hif1a antibody in lab because of the extensive process of validation that CST performs on all of their antibodies, and the ability to order a bunch from the same lot at once.

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 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

Very clean results with this antibody. Minimal background on film and with chemiluminescent imager. Was able to sufficiently demonstrate knockdown of Hif1a among samples incubated in 100 uM DFO for 6 hours. I have found it best to use a 1:1 ratio of ECL to FEMTO in order to fully bring out the antibody on film.

Additional Notes

None

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Summary

The Good

Clean and reliable

The Bad

Requires FEMTO for adequate exposure

The Bottom Line

A top-notch product that I have used for years and will continue to use for years to come.

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