BD Biosciences
Purified Mouse Anti-Human HIF-1α
610958
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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.
Western Blot
A549 cell lysate exposed to normoxia (21% O2) and hypoxia (1% O2)
1:1000 in 5% skimmed milk for overnight at 4 degrees
5% Skimmed milk
Rabbit anti mouse (biorad)- 1:10,000 in skimmed milk 5% skimmed milk for 1 hr at RT
N/A
ECL (Thermo)
A very specific single band was observed around 120KD in hypoxic sample.
Specific band
None
Ideal antibody for western. Very specific.