Human/Mouse/Rat HIF-1 alpha/HIF1A Antibody

Detection of Human HIF‑1 alpha /HIF1A by Western Blot. Western blot shows lysates of MCF-7 human breast cancer cell line untreated (-) or treated (+) with 150 µM CoCl₂for 16 hours. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human/Mouse/Rat HIF-1a/HIF1A Antigen-affinity Purified Polyclonal Antibody, followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for HIF-1a/HIF1A at approximately 120 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of HIF‑1 alpha /HIF1A-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-1a/DNA complexes were immunoprecipitated using 5 µg Goat Anti-Human/Mouse/Rat HIF-1a/HIF1A 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.

HIF‑1 alpha /HIF1A Specificity is Shown by Immunocytochemistry in Knockout Cell Line. HIF-1a/HIF1A was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line treated with DFO but is not detected in HIF-1a/HIF1A knockout (KO) HeLa Human Cell Line cell line using Goat Anti-Human/Mouse/Rat HIF-1a/HIF1A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1935) at 0.3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Human HIF‑1 alpha /HIF1A by Simple Western^TM. 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-1a/HIF1A at approximately 115 kDa (as indicated) using 5 µg/mL of Goat Anti-Human/Mouse/Rat HIF-1a/HIF1A 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.