Cobalt Chloride treated HeLa cells were subjected to SDS PAGE followed by western blot with FNab09940(HIF1a Antibody) at dilution of 1:5000
Immunofluorescent analysis of ( -20°C Ethanol) fixed Cobalt Chloride treated HeLa cells using FNab09940 (HIF1a antibody) at dilution of 1:50 and Alexa Fluor 488-Conjugated Goat Anti-Mouse IgG(H+L)
Background
Under normal oxygen conditions, HIF1a is continuously produced and destroyed, in a process involving hydroxylation, interaction with von Hippel-Lindau (VHL) protein, polyubiquitylation and subsequent proteasomal degradation. Under hypoxic conditions, hydroxylation is impaired and HIF1a is stabilized. HIF1a localizes in cytoplasm in normoxia, but it can translocate into nuclear in response to hypoxia. The calculated molecular weight of HIF1a is 93 kDa, but the modified protein HIF1a is about 110-120kDa (PMID: 11698256, .PMID: 7539918).