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.
Human HIF-1 alpha/HIF1A Antibody
R&D Systems | Catalog # MAB19352
Recombinant Monoclonal Antibody.
Key Product Details
Validated by
Knockout/Knockdown
Species Reactivity
Human
Applications
Knockout Validated, Immunohistochemistry, Western Blot, Immunocytochemistry, Simple Western
Label
Unconjugated
Antibody Source
Recombinant Monoclonal Rabbit IgG Clone # 2443C
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Product Specifications
Immunogen
E. coli-derived recombinant human HIF-1 alpha/HIF1A
Arg575-Asn826
Accession # Q16665.1
Arg575-Asn826
Accession # Q16665.1
Specificity
Detects human HIF-1 alpha/HIF1A in direct ELISAs.
Clonality
Monoclonal
Host
Rabbit
Isotype
IgG
Scientific Data Images for 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 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.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.Applications for Human HIF-1 alpha/HIF1A Antibody
Application
Recommended Usage
Immunocytochemistry
1-25 µg/mL
Sample: Immersion fixed MCF-7 human breast cancer cell line and HeLa human cervical epithelial carcinoma cell line treated with DFO and HeLa knockouts treated with DFO
Sample: Immersion fixed MCF-7 human breast cancer cell line and HeLa human cervical epithelial carcinoma cell line treated with DFO and HeLa knockouts treated with DFO
Immunohistochemistry
5-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human liver
Sample: Immersion fixed paraffin-embedded sections of human liver
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.
Simple Western
20 µg/mL
Sample: HepG2 human hepatocellular carcinoma cell line treated with DFO
Sample: HepG2 human hepatocellular carcinoma cell line treated with DFO
Western Blot
2 µg/mL
Sample: HIF-1 alpha/HIF1A human hepatocellular carcinoma cell line treated with DFO
Sample: HIF-1 alpha/HIF1A human hepatocellular carcinoma cell line treated with DFO
Formulation, Preparation, and Storage
Purification
Protein A or G purified from cell culture supernatant
Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
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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.
Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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.
Calculators
Background: HIF-1 alpha/HIF1A
Long Name
Hypoxia Inducible Factor 1 Subunit Alpha
Alternate Names
BHLHE78, HIF 1A, HIF-1a, HIF1 alpha, HIF1A, MOP1, PASD8
Gene Symbol
HIF1A
UniProt
Additional HIF-1 alpha/HIF1A Products
Product Documents for Human HIF-1 alpha/HIF1A Antibody
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human HIF-1 alpha/HIF1A Antibody
For research use only
Citations for Human HIF-1 alpha/HIF1A Antibody
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Antigen Retrieval Protocol (PIER)
- Antigen Retrieval for Frozen Sections Protocol
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- Chromogenic IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Chromogenic Immunohistochemistry Staining of Frozen Tissue
- ClariTSA™ Fluorophore Kits
- Detection & Visualization of Antibody Binding
- Fluorescent IHC Staining of Frozen Tissue Protocol
- Graphic Protocol for Heat-induced Epitope Retrieval
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Graphic Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- ICC Cell Smear Protocol for Suspension Cells
- ICC Immunocytochemistry Protocol Videos
- ICC for Adherent Cells
- IHC Sample Preparation (Frozen sections vs Paraffin)
- Immunocytochemistry (ICC) Protocol
- Immunocytochemistry Troubleshooting
- Immunofluorescence of Organoids Embedded in Cultrex Basement Membrane Extract
- Immunofluorescent IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Immunohistochemistry Frozen Troubleshooting
- Immunohistochemistry Paraffin Troubleshooting
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Protocol for Heat-Induced Epitope Retrieval (HIER)
- Protocol for Making a 4% Formaldehyde Solution in PBS
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Fluorescent ICC Staining of Cell Smears - Graphic
- Protocol for the Fluorescent ICC Staining of Cultured Cells on Coverslips - Graphic
- Protocol for the Preparation & Fixation of Cells on Coverslips
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections - Graphic
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections - Graphic
- Protocol for the Preparation and Fluorescent ICC Staining of Cells on Coverslips
- Protocol for the Preparation and Fluorescent ICC Staining of Non-adherent Cells
- Protocol for the Preparation and Fluorescent ICC Staining of Stem Cells on Coverslips
- Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- Protocol for the Preparation of a Cell Smear for Non-adherent Cell ICC - Graphic
- R&D Systems Quality Control Western Blot Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: Immunohistochemistry
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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