Human/Rat HIF-2 alpha/EPAS1 Antibody

Detection of Human and Rat HIF-2 alpha/EPAS1 by Western Blot.

Western blot shows lysates of MCF-7 human breast cancer cell line and PC-12 rat adrenal pheochromocytoma cell line untreated (-) or treated (+) with 150 µM CoCl₂for 16 h. PVDF membrane was probed with 1 µg/mL of Human/Rat HIF-2a/EPAS1 Polyclonal Antibody, followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for HIF-2a/EPAS1 at approximately 115 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of HIF‑2 alpha/EPAS1-regulated Genes by Chromatin Immunoprecipitation.

BG01V human embryonic stem cells treated with 150 µM CoCl2 overnight were fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. HIF-2a/EPAS1/DNA complexes were immunoprecipitated using 5 µg Goat Anti-Human/Rat HIF-2a/EPAS1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2886) or control antibody (Catalog # AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (Catalog # BAF109). Immunocomplexes were captured using 50 µL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. Theoct-3/4promoter was detected by standard PCR.

Detection of Human HIF-2 alpha/EPAS1 by Western Blot

The DU145FP cells express higher levels of cancer stemness related genes. (A) Transcriptome analyses of prostate cancer stemness-related markers. The Log2 transformed ratio of FPKM values (e.g., FP/WT) are indicated by color-coded index bars. (B) Cells were stained with fluorophore conjugated monoclonal antibodies against CD44 and EPCAM, the antibody against integrin alpha2 beta3 without fluorophore conjugation was detected by using secondary antibody conjugated with fluorophore, and stained cells were analyzed by flow cytometry. The scatter plots and histograms show representative results. CTRL denote cell stained with isotype control antibody or only secondary antibody; ST denote cell stained with antibody against EPCAM and integrin alpha2 beta3; US denote unstained cells. (C) Immunoblotting confirmation of the protein expression of A-tubulin, GAPDH, SOX9, SLUG, HIF1A, HIF2A, CD44 and integrin alpha2 beta3. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28698547), licensed under a CC-BY license. Not internally tested by R&D Systems.