HIF-2 alpha, also known as hypoxia-inducible factor 2, endothelial PAS domain protein-1, and member of PAS superfamily 2 is part of the HIF family of proteins. The HIF family is composed of HIF-1, HIF-2 and HIF-3, where HIF-2 is a dimeric protein that consists of an alpha and beta subunit.HIF-2 alphais activated in the presence of oxygen deprivation, or hypoxia, via prolyl hydroxylase-domain enzymes (PHDs) that facilitate its degradation. HIF-2 alphais expressed in endothelial and intestinal cells, as well as in tumor tissue of a variety of prominent cancers, and is not typically detected in normoxic conditions. Given the fact that loss of HIF-1 alpha was a notable hallmark of renal cell carcinoma, HIF-2 alpha and the hypoxia mediated gene therapy has been an area of interest in research renal cell carcinoma therapy. Using a HIF-2 alpha antibody in multiple research applications allowed the following scientific groups a better understand of the role of hypoxia and clear cell renal cell carcinoma.
HIF-2 alpha/EPAS1 Antibody [NB100-122] - Simple Western lane view shows a specific band for HIF-2 alpha in 0.5 mg/ml of Hypoxic HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system
First, Emerling et al used a HIF-2 alpha antibody to help better understand how the potential HIF-2 alphatarget gene, CDCP1, might be associated with survival in clear cell renal cell carcinoma (ccRCC). While investigating the effects of survival in hypoxic conditions, Emerling’s group noted that hypoxia caused a marked increase of CDCP1. With this information, HIF-2 alpha antibody was used in western blot on immortalized human mammary epithelial cell lines with retroviral shRNA CDCP1 knockdown features. MCF10A cells were created that expressed shRNAs targeting HIF-1 alpha, HIF-2 alphaand ARNT. A HIF-2 alpha antibody was used and showed that the knockdown of HIF-2 alpha was indeed successful. Additionally, a HIF-2 alpha antibody was used in a co-immunoprecipitation experiment to show that HIF-2 alphabound to the CDCP1 promoter in the presence of a hypoxic environment. Lastly, a HIF-2 alpha antibody was used once again in western blot to show that VHL-deficient RCC cell lines express high CDCP1 protein levels alongside HIF-2 alphaexpression, suggesting that CDCP1 is a probable target for ccRCC therapy.
Next, Qui et al used a HIF-2 alpha antibody in their study on HIF-2 alphadependent lipid storage and its effect on endoplasmic reticulum homeostasis in clear cell renal cell carcinoma (ccRCC). It was preliminarily established that HIF-2 alphadependent PLIN2 gene expression was associated with lipid storage, cellular proliferation, and cellular viability. To investigate whether HIF activation promotes PLIN2 expression, transcriptome profiling of tumors that were negative for HIF-1 alpha, HIF-1 alpha and HIF-2 alphaexpressing, and HIF-2 alphaexpressing only was first performed. In western blot, a HIF-2 alpha antibody showed that HIF-2 alphatargets in ccRCC had a similar expression pattern, which suggest that PLIN2 levels increased early in ccRCC disease progression and correlate with elevated HIF-2 alphaactivity.
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