Great Antibody for HIF-1a Western Blot

Surgery
University of California, San Diego
Postdoctoral Research Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

BD Biosciences

Product Name:

Purified Mouse Anti-Human HIF-1α

Catalog Number:

610958

The main focus of my research is understanding the interplay of hypoxia in non-small cell lung cancer. Hypoxia is low oxygen conditions, which is generally developed in different kinds of tumors because of imbalance of oxygen supply and demand. Hypoxia inducible factor (HIF) are transcription factors which are activated during hypoxia. HIF family members includes- HIF-1α, HIF-1β, HIF-2α, HIF-2β, HIF-3α and HIF-3β. During normal oxygen conditions (normoxia), the alpha units of HIF are hydroxylated at proline residue, which is recognized by ubiquitin ligase, leading to their proteosomal degradation. Hence, HIFα are not stabilized and degraded during normoxia. But in hypoxic conditions, HIF prolyl hydroxylase are inhibited, hence HIF α are stabilized. HIF-1α targets and regulate gene-expression of multiple other genes during hypoxia, to promote cell survival in low oxygen conditions. HIF-1α is routinely used as positive control/marker for hypoxia.

Experimental Design and Results Summary

Applications

Western Blot

Sample

A549 cell lysate exposed to normoxia (21% O2) and hypoxia (1% O2)

Primary Incubation

1:1000 in 5% skimmed milk for overnight at 4 degrees

Blocking Agent

5% Skimmed milk

Secondary Incubation

Rabbit anti mouse (biorad)- 1:10,000 in skimmed milk 5% skimmed milk for 1 hr at RT

Tertiary Incubation

N/A

Detection

ECL (Thermo)

Results Summary

A very specific single band was observed around 120KD in hypoxic sample.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

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Summary

The Good

Specific band

The Bad

None

The Bottom Line

Ideal antibody for western. Very specific.

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