Human HIF-1 alpha/HIF1A Antibody

Name: Human HIF-1 alpha/HIF1A Antibody
Brand: R&D Systems
SKU: MAB19352
Price: 134 USD

Catalog # MAB19352 | R&D Systems, Inc. a Bio-Techne Brand

Product Datasheet / COA / SDS

Recombinant Monoclonal Antibody.

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Product Summary
Scientific Data
Applications
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Human

Applications

Immunocytochemistry, Immunohistochemistry, Knockout Validated, Simple Western, Western Blot

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # 2443C

Product Summary for Human HIF-1 alpha/HIF1A Antibody

Immunogen

E. coli-derived recombinant human HIF-1 alpha/HIF1A
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 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.

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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

Immunohistochemistry

5-25 µg/mL
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

Western Blot

2 µg/mL
Sample: HIF-1 alpha/HIF1A human hepatocellular carcinoma cell line treated with DFO

Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation and Storage

Purification

Protein A or G purified from cell culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

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

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

Alternate Names

BHLHE78, HIF 1A, HIF-1a, HIF1 alpha, HIF1A, MOP1, PASD8

Entrez Gene IDs

3091 (Human); 15251 (Mouse); 29560 (Rat)

Gene Symbol

HIF1A

UniProt

Q16665.1

Product Documents for Human HIF-1 alpha/HIF1A Antibody

Product Datasheets

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COA

SDS

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Product Specific Notices for Human HIF-1 alpha/HIF1A Antibody

For research use only

FAQs for Human HIF-1 alpha/HIF1A Antibody

Showing 1 - 2 of 2 FAQs Showing All

Q: Which antibody(ies) do you recommend for the detection of HIF-1a by immunohistochemistry in the sections of paraffin-embedded mouse liver samples? I would appreciate if you can give me several choices and rank them in the order of performance. My goal is to distinguish HIF upregulation by prolyl hydroxylase inhibitor in different liver cells.

A: All of our antibodies are of high quality and are well tested/validated in species/applications we list on the datasheet. However, we suggest the following four HIF-1 alpha antibodies based upon customer reviews, as well as the number of peer reviewed publications in which these products have been cited by researchers from reputed institutes. (1) HIF-1 alpha Antibody (H1alpha67) (cat# NB100-105) (cited in at least 218 peer reviewed publications) (2) HIF-1 alpha Antibody (cat# NB100-479) (cited in at least 51 peer reviewed publications) (3) HIF-1 alpha Antibody (H1alpha67) (cat# NB100-123 ) (cited in at least 38 peer reviewed publications) (4) HIF-1 alpha Antibody (cat# NB100-449) (cited in at least 31 peer reviewed publications).
Q: What is the molecular weight (kDa) of protein HIF 1 alpha in western blot?

A: The theoretical molecular weight of HIF 1-alpha is ~93kDa. However, you will likely see a band between 100-120kDa due to phosphorylation.