HIF-1α (D1S7W) XP^® Rabbit mAb #36169

Filter:

WB
IP
IF
F
ChIP
C&R

Supporting Data

REACTIVITY	H M Mk
SENSITIVITY	Endogenous
MW (kDa)	120
Source/Isotype	Rabbit IgG

Application Key:

WB-Western Blotting
IP-Immunoprecipitation
IF-Immunofluorescence
F-Flow Cytometry
ChIP-Chromatin Immunoprecipitation
C&R-CUT & RUN

Species Cross-Reactivity Key:

H-Human
M-Mouse
Mk-Monkey

Product Information

Product Usage Information

For optimal ChIP and ChIP-seq results, use 5 μl of antibody and 10 μg of chromatin (approximately 4 x 10⁶ cells) per IP. This antibody has been validated using SimpleChIP^® Enzymatic Chromatin IP Kits.

The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.

Application	Dilution
Western Blotting	1:1000
Immunoprecipitation	1:50
Immunofluorescence (Immunocytochemistry)	1:400 - 1:1600
Flow Cytometry (Fixed/Permeabilized)	1:100 - 1:400
Chromatin IP	1:100
Chromatin IP-seq	1:100
CUT&RUN	1:100

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #36199.

Protocol

Specificity / Sensitivity

HIF-1α (D1S7W) XP^® Rabbit mAb recognizes endogenous levels of total HIF-1α protein. This antibody does not cross-react with HIF-2α protein.

Species Reactivity:

Human, Mouse, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu478 of human HIF-1α protein.

Background

Hypoxia-inducible factor 1 (HIF1) is a heterodimeric transcription factor that plays a critical role in the cellular response to hypoxia (1). The HIF1 complex consists of two subunits, HIF-1α and HIF-1β, which are basic helix-loop-helix proteins of the PAS (Per, ARNT, Sim) family (2). HIF1 regulates the transcription of a broad range of genes that facilitate responses to the hypoxic environment, including genes regulating angiogenesis, erythropoiesis, cell cycle, metabolism, and apoptosis. The widely expressed HIF-1α is typically degraded rapidly in normoxic cells by the ubiquitin/proteasomal pathway. Under normoxic conditions, HIF-1α is proline hydroxylated leading to a conformational change that promotes binding to the von Hippel-Lindau protein (VHL) E3 ligase complex; ubiquitination and proteasomal degradation follows (3,4). Both hypoxic conditions and chemical hydroxylase inhibitors (such as desferrioxamine and cobalt) inhibit HIF-1α degradation and lead to its stabilization. In addition, HIF-1α can be induced in an oxygen-independent manner by various cytokines through the PI3K-AKT-mTOR pathway (5-7).

HIF-1β is also known as AhR nuclear translocator (ARNT) due to its ability to partner with the aryl hydrocarbon receptor (AhR) to form a heterodimeric transcription factor complex (8). Together with AhR, HIF-1β plays an important role in xenobiotics metabolism (8). In addition, a chromosomal translocation leading to a TEL-ARNT fusion protein is associated with acute myeloblastic leukemia (9). Studies also found that ARNT/HIF-1β expression levels decrease significantly in pancreatic islets from patients with type 2 diabetes, suggesting that HIF-1β plays an important role in pancreatic β-cell function (10).

Database Links

UniProt ID: Q16665

Entrez-Gene Id: 3091

Limited Uses

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For Research Use Only. Not For Use In Diagnostic Procedures.

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HIF-1α (D1S7W) XP® Rabbit mAb #36169