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Hypoxia-inducible factor 1 alpha (HIF-1 alpha) is a subunit of a heterodimeric transcription factor hypoxia-inducible factor 1 (HIF-1) that is encoded by the HIF1A gene.
HIF-1 alpha can be a challenging target to work with in western blot.
HIF-1 alpha is only stabilized at O2 concentrations below 5%. Under normoxic conditions, HIF-1 alpha has a short half-life and may be degraded within 5-8 minutes in both nuclear and cytoplasmic compartments. Therefore, proper sample preparation is critical for western blot success. If care hasn’t been taken with sample preparation, you might not see any bands on western blot.
Note: the observed band size of HIF-1 alpha may not be the same as predicted in western blot due to the different forms of HIF-1 alpha, including:
Negative control: Most normal tissues or cells (other than kidney or heart) should work as a negative control, as HIF-1 alpha will degrade when taken out of hypoxic conditions.