Western Blot: HIF-1 alpha [Hydroxy Pro564] Antibody [NB110-74679] HIF-1alpha HYDROXYP564 antibody shows detection (left panel) of hydroxylated HIF-1alpha in nuclear extracts of A549 cells treated with MG132 (a proteosome inhibitor). Hydroxyproline is not recognized on HIF-1alpha when cells are first treated with DFO, a prolyl hydroxylase inhibitor that prevents HIF hydroxylation. Control staining is shown (right panel) using conventional anti-HIF-1alpha. The asterisk marks a band approximately 110 kDa in size corresponding to HIF1-alpha. The primary antibody was used at a 1:1,000 dilution in 2% BLOTTO.