Human HIF-1 alpha/HIF1A Antibody

Detection of Human HIF-1 alpha/HIF1A by Western Blot. Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line untreated (-) or treated (+) with 1 mM DFO for overnight. PVDF membrane was probed with 2 µg/mL of Rabbit Anti-Human HIF-1 alpha/HIF1A Monoclonal Antibody (Catalog # MAB19352) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for HIF-1 alpha/HIF1A at approximately 120 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

HIF-1 alpha/HIF1A in MCF-7 Human Cell Line. HIF-1 alpha/HIF1A was detected in immersion fixed MCF-7 human breast cancer cell line using Rabbit Anti-Human HIF-1 alpha/HIF1A Monoclonal Antibody (Catalog # MAB19352) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

HIF-1 alpha/HIF1A in HeLa Human Cell Line. HIF-1 alpha/HIF1A was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line treated with DFO (left panel, positive stain) or HeLa knockouts treated with DFO (right panel, negative stain) using Rabbit Anti-Human HIF-1 alpha/HIF1A Monoclonal Antibody (Catalog # MAB19352) at 1 µg/mL for 3 hours at room temperature. Cells were stained using a Goat anti-Rabbit IgG Secondary Antibody, Alexa Fluor 488 (green) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

HIF-1 alpha/HIF1A in Human Liver. HIF-1 alpha/HIF1A was detected in immersion fixed paraffin-embedded sections of human liver using Rabbit Anti-Human HIF-1 alpha/HIF1A Monoclonal Antibody (Catalog # MAB19352) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and nuclei in hepatocytes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of Human HIF-1 alpha/HIF1A by Simple Western^TM. Simple Western lane view shows lysates of HepG2 human hepatocellular carcinoma cell line untreated (-) or treated (+) with 1 mM DFO for overnight, loaded at 0.2 mg/mL. A specific band was detected for HIF-1 alpha/HIF1A at approximately 116 kDa (as indicated) using 20 µg/mL of Rabbit Anti-Human HIF-1 alpha/HIF1A Monoclonal Antibody (Catalog # MAB19352). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Western Blot Shows Human HIF-1 alpha/HIF1A Specificity by Using Knockout Cell Line. Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and HIF-1 alpha/HIF1A knockout HeLa cell line (KO) untreated (-) or treated (+) with 1 mM DFO for overnight. PVDF membrane was probed with 2 µg/mL of Rabbit Anti-Human HIF-1 alpha/HIF1A Monoclonal Antibody (Catalog # MAB19352) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for HIF-1 alpha/HIF1A at approximately 120 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.