Human HIF-1 alpha/HIF1A Antibody

Recombinant Monoclonal Antibody
Catalog # Availability Size / Price Qty
MAB19352-100
MAB19352-SP
Detection of Human HIF-1 alpha/HIF1A by Western Blot.
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Human HIF-1 alpha/HIF1A Antibody Summary

Species Reactivity
Human
Specificity
Detects human HIF-1 alpha/HIF1A in direct ELISAs.
Source
Recombinant Monoclonal Rabbit IgG Clone # 2443C
Purification
Protein A or G purified from cell culture supernatant
Immunogen
E. coli-derived recombinant human HIF-1 alpha/HIF1A
Arg575-Asn826
Accession # Q16665.1
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
2 µg/mL
See below
Simple Western
20 µg/mL
See below
Immunohistochemistry
5-25 µg/mL
See below
Immunocytochemistry
1-25 µg/mL
See below
Knockout Validated
HIF-1 alpha/HIF1A is specifically detected in HeLa human cervical epithelial carcinoma parental cell line but is not detectable in HIF-1 alpha/HIF1A knockout HeLa cell line.
 

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Human HIF-1 alpha/HIF1A antibody by Western Blot. View Larger

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.

Immunocytochemistry HIF-1 alpha/HIF1A antibody in MCF-7 Human Cell Line by Immunocytochemistry (ICC). View Larger

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.

Immunocytochemistry HIF-1 alpha/HIF1A antibody in HeLa Human Cell Line by Immunocytochemistry (ICC). View Larger

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.

Immunohistochemistry HIF-1 alpha/HIF1A antibody in Human Liver by Immunohistochemistry (IHC-P). View Larger

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.

Simple Western Detection of Human HIF-1 alpha/HIF1A antibody by Simple Western<sup>TM</sup>. View Larger

Detection of Human HIF-1 alpha/HIF1A by Simple WesternTM. 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.

Knockout Validated Western Blot Shows Human HIF-1 alpha/HIF1A Antibody Specificity by Using Knockout Cell Line. View Larger

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.

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
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Shipping
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/HIF1A

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.

Long Name
Hypoxia Inducible Factor 1 Subunit Alpha
Entrez Gene IDs
3091 (Human); 15251 (Mouse); 29560 (Rat)
Alternate Names
AINT; anti-HIF-1 alpha; anti-HIF1A; ARNT interacting protein; ARNT-interacting protein; Basic-helix-loop-helix-PAS protein MOP1; BHLHE78; Class E basic helix-loop-helix protein 78; HIF 1A; HIF1 alpha; HIF-1 alpha; HIF1; HIF1A; HIF-1a; HIF-1alpha; HIF-1-alpha; HIF1-alpha; hypoxia inducible factor 1 alpha subunit, hypoxia inducible factor 1 subunit alpha; hypoxia inducible factor 1, alpha subunit (basic helix-loop-helix transcription factor); hypoxia-inducible factor 1-alpha; Member of PAS protein 1; member of PAS superfamily 1; MOP1; PAS domain-containing protein 8; PASD8

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