Western blot shows lysates of MCF‑7 human breast cancer cell line untreated (-) or treated (+) with 150 μM CoCl2 for 16 hours. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human/Mouse/Rat HIF‑1 alpha Antigen-affinity Purified Polyclonal Antibody, followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for HIF‑1 alpha at approximately 120 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of HIF‑1 alpha -regulated Genes by Chromatin Immunoprecipitation.
Mouse primary kidney cells treated with 150 μM CoCl2 for overnight were fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. HIF‑1 alpha /DNA complexes were immunoprecipitated using 5 μg Goat Anti-Human/Mouse/Rat HIF‑1 alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1935) 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 epo promoter was detected by standard PCR.
Detection of Human HIF‑1 alpha by Simple WesternTM.
Simple Western lane view shows lysates of A549 human lung carcinoma cell line untreated (-) or treated (+) with Hypoxia (1% O2), loaded at 0.2 mg/mL. A specific band was detected for HIF‑1 alpha at approximately 115 kDa (as indicated) using 5 µg/mL of Goat Anti-Human/Mouse/Rat HIF‑1 alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1935) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
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-1 alpha
The hypoxia-inducible transcription factor 1 alpha (HIF-1 alpha ) is the regulated member of the transcription factor heterodimer HIF-1. HIF-1 binds to hypoxia-response elements (HREs) in the promoters of many genes involved in adapting to an environment of insufficient oxygen or hypoxia. Hypoxic tissue environments occur in vascular and pulmonary diseases as well as cancer, which illustrates the broad impact of gene regulation by HIF-1 alpha.
Hypoxia-inducible Transcription Factor 1 alpha
Entrez Gene IDs:
3091 (Human); 15251 (Mouse); 29560 (Rat)
ARNT-interacting protein; Basic-helix-loop-helix-PAS protein MOP1; BHLHE78; Class E basic helix-loop-helix protein 78; HIF1 alpha; HIF-1 alpha; HIF1A; HIF-1-alpha; HIF1-alpha; hypoxia inducible factor 1, alpha subunit (basic helix-loop-helix transcriptionfactor); hypoxia-inducible factor 1-alpha; Member of PAS protein 1; member of PAS superfamily 1; MOP1HIF1-ALPHA; PAS domain-containing protein 8; PASD8alpha subunit (basic helix-loop-helix transcriptionfactor)
R&D Systems personnel manually curate a database that contains references using R&D Systems products.
The data collected includes not only links to publications in PubMed,
but also provides information about sample types, species, and experimental conditions.
The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.