Figure 1: Treatment of HeLa cells with the hydroxylase inhibitor dimethyloxaloylglycine (DMOG) results in decreased hydroxylation of HIF-1α, as detected by the PathScan® Hydroxy-HIF-1α (Pro564) Sandwich ELISA Kit #13201, but does not affect the level of total HIF-1α detected by PathScan® Total HIF-1α Sandwich ELISA Kit #13127. Absorbance at 450 nm is shown in the top figures while corresponding western blots using a total HIF-1α antibody (left) and Hydroxy-HIF-1α (Pro564) (D43B5) XP® Rabbit mAb #3434 (right) are shown in the bottom figures. Treatment of HeLa cells with the proteasome inhibitor MG-132 #2194 stabilizes HIF-1α protein.
Figure 2: The relationship between protein concentration of lysates from HeLa cells, untreated or treated with MG-132 #2194, and the absorbance 450 nm as detected by the PathScan® Hydroxy-HIF-1α (Pro564) Sandwich ELISA Kit is shown.