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 CoCl2 for 16 h. PVDF membrane was probed with 1 µg/mL of Human/Rat HIF‑2 alpha /EPAS1 Polyclonal Antibody, followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for HIF‑2 alpha /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‑2 alpha /EPAS1/DNA complexes were immunoprecipitated using 5 μg Goat Anti-Human/Rat HIF‑2 alpha /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. The oct-3/4 promoter was detected by standard PCR.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: HIF-2 alpha/EPAS1
The hypoxia-inducible transcription factor 2 alpha (HIF-2 alpha ) is stabilized in hypoxic tissue and, similarly to HIF-1 alpha, complexes with Aryl hydrocarbon receptor nuclear translocator (ARNT). Both the HIF-1 and HIF-2 complexes bind hypoxia-response elements (HREs) in the promoters of many genes involved in adapting to an environment of insufficient oxygen or hypoxia. HIF-1 and HIF-2 do not appear completely redundant, although specific functions are only beginning to be elucidated. Hypoxic tissue environments occur in vascular and pulmonary diseases as well as cancer, which illustrates the potentially broad impact of gene regulation by both HIF-1 alpha and HIF-2 alpha.
Hypoxia-inducible Transcription Factor 2 alpha
Entrez Gene IDs:
Basic-helix-loop-helix-PAS protein MOP2; BHLHE73; bHLHe73ECYT4; Class E basic helix-loop-helix protein 73; endothelial PAS domain protein 1; endothelial PAS domain-containing protein 1; EPAS1; EPAS-1; HIF 2A; HIF-1alpha-like factor; HIF2 alpha; HIF-2 alpha; hif2a angiogenesis; HIF2A; HIF2AHIF-2-alpha; HLF; HLFHIF-1-alpha-like factor; hypoxia-inducible factor 2 alpha; Hypoxia-inducible factor 2-alpha; Member of PAS protein 2; MOP2; MOP2HIF2-alpha; PASD2PAS domain-containing protein 2
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