Recently, we at Novus Biologicals have added a new batch of monoclonal anti-HIF-1 alpha antibody to our hypoxia catalogue, developed from H1alpha67 hybridoma cultures. This furthers the role that HIF-1 antibodies play in cancer research.
HIF-1 becomes active under hypoxic conditions. Its function is the transcription or blocking of a number of genes, to preserve cell viability at times of low oxygen stress. However, HIF transcription can also be activated in non-hypoxic conditions, and HIF expression has been shown to be a factor in tumour development. Therefore both normal and cancerous cell-lines are used in HIF-1 antibody studies.
HIF-1 is a heterodimer complex composed of identical β and α subunits, which must both be expressed for HIF-1 to become active. Under normoxic conditions, the α-subunit is degraded by the cellular proteasome, a process that becomes inactive if hypoxia occurs. The β-subunit is continually expressed.
-Western%20Blot-NB100-105-img0005.jpg "Western Blot: HIF-1 alpha Antibody")
Western Blot: HIF-1 alpha Antibody
HIF-α has three isoforms, known to be regulated by prolyl hydroxylation of the O2-dependent degradation domain, during the posttranslational phase. Prolyl hydroxylases target the protein via interaction with VHL (von Hippel-Lindau), one of the components of the E3 ubiquitin-ligase complex (VBC). VBC links ubiquitin to HIFα, tagging it for degradation by the cellular proteolytic proteasomal complex.
Additional hydroxylation takes place on the HIF1α and HIF2α C-terminus. This inhibits the binding of co-activators such as p300 and CREB-binding protein (CBP), thus inhibiting HIF-1 activation. The entire hydroxylation pathway is catalysed by 3-PHD (prolyl hydroxylase domain) isoforms, plus FIH (factor inhibiting HIF) proteins. Their activity is dependent upon cofactor Fe2+ , 2- oxoglutarate and oxygen.
Over-expression of PHD-3 is associated with aggressive pancreatic tumours, while p300/CREB studies have shown similar results. Therefore HIFα antibody studies continue to be important in cancer research.
Novus Biologicals offers many HIF-1 alpha reagents for your research needs including:
