ARG41701

anti-HIF-1 alpha antibody

anti-HIF-1 alpha antibody for Flow cytometry,ICC/IF,IHC-Formalin-fixed paraffin-embedded sections,Immunoprecipitation,Western blot and Human

Overview

Product Description Rabbit Polyclonal antibody recognizes HIF-1 alpha
Tested Reactivity Hu
Tested Application FACS, ICC/IF, IHC-P, IP, WB
Host Rabbit
Clonality Polyclonal
Isotype IgG
Target Name HIF-1 alpha
Antigen Species Human
Immunogen Synthetic peptide of Human HIF-1 alpha.
Conjugation Un-conjugated
Alternate Names Class E basic helix-loop-helix protein 78; Basic-helix-loop-helix-PAS protein MOP1; Hypoxia-inducible factor 1-alpha; PAS domain-containing protein 8; HIF1-alpha; HIF1-ALPHA; HIF1; MOP1; HIF-1-alpha; PASD8; HIF-1A; HIF-1alpha; Member of PAS protein 1; ARNT-interacting protein; bHLHe78

Application Instructions

Application Suggestion
Tested Application Dilution
FACS1:30
ICC/IF1:50 - 1:200
IHC-P1:50 - 1:200
IP1:30
WB1:500 - 1:2000
Application Note * The dilutions indicate recommended starting dilutions and the optimal dilutions or concentrations should be determined by the scientist.
Observed Size 92-130 kDa

Properties

Form Liquid
Purification Affinity purified.
Buffer PBS (pH 7.4), 150 mM NaCl, 0.02% Sodium azide and 50% Glycerol.
Preservative 0.02% Sodium azide
Stabilizer 50% Glycerol
Storage Instruction For continuous use, store undiluted antibody at 2-8°C for up to a week. For long-term storage, aliquot and store at -20°C. Storage in frost free freezers is not recommended. Avoid repeated freeze/thaw cycles. Suggest spin the vial prior to opening. The antibody solution should be gently mixed before use.
Note For laboratory research only, not for drug, diagnostic or other use.

Bioinformation

Database Links

GeneID: 3091 Human HIF1A

Swiss-port # Q16665 Human Hypoxia-inducible factor 1-alpha

Gene Symbol HIF1A
Gene Full Name hypoxia inducible factor 1, alpha subunit (basic helix-loop-helix transcription factor)
Background This gene encodes the alpha subunit of transcription factor hypoxia-inducible factor-1 (HIF-1), which is a heterodimer composed of an alpha and a beta subunit. HIF-1 functions as a master regulator of cellular and systemic homeostatic response to hypoxia by activating transcription of many genes, including those involved in energy metabolism, angiogenesis, apoptosis, and other genes whose protein products increase oxygen delivery or facilitate metabolic adaptation to hypoxia. HIF-1 thus plays an essential role in embryonic vascularization, tumor angiogenesis and pathophysiology of ischemic disease. Alternatively spliced transcript variants encoding different isoforms have been identified for this gene. [provided by RefSeq, Jul 2011]
Function Functions as a master transcriptional regulator of the adaptive response to hypoxia. Under hypoxic conditions, activates the transcription of over 40 genes, including erythropoietin, glucose transporters, glycolytic enzymes, vascular endothelial growth factor, HILPDA, and other genes whose protein products increase oxygen delivery or facilitate metabolic adaptation to hypoxia. Plays an essential role in embryonic vascularization, tumor angiogenesis and pathophysiology of ischemic disease. Binds to core DNA sequence 5'-[AG]CGTG-3' within the hypoxia response element (HRE) of target gene promoters. Activation requires recruitment of transcriptional coactivators such as CREBPB and EP300. Activity is enhanced by interaction with both, NCOA1 or NCOA2. Interaction with redox regulatory protein APEX seems to activate CTAD and potentiates activation by NCOA1 and CREBBP. Involved in the axonal distribution and transport of mitochondria in neurons during hypoxia. [UniProt]
Cellular Localization Cytoplasm. Nucleus. Nucleus speckle. Note=Colocalizes with HIF3A in the nucleus and speckles (By similarity). Cytoplasmic in normoxia, nuclear translocation in response to hypoxia (PubMed:9822602). [UniProt]
Calculated MW 93 kDa
PTM In normoxia, is hydroxylated on Pro-402 and Pro-564 in the oxygen-dependent degradation domain (ODD) by EGLN1/PHD2 and EGLN2/PHD1. EGLN3/PHD3 has also been shown to hydroxylate Pro-564. The hydroxylated prolines promote interaction with VHL, initiating rapid ubiquitination and subsequent proteasomal degradation. Deubiquitinated by USP20. Under hypoxia, proline hydroxylation is impaired and ubiquitination is attenuated, resulting in stabilization.

In normoxia, is hydroxylated on Asn-803 by HIF1AN, thus abrogating interaction with CREBBP and EP300 and preventing transcriptional activation. This hydroxylation is inhibited by the Cu/Zn-chelator, Clioquinol.

S-nitrosylation of Cys-800 may be responsible for increased recruitment of p300 coactivator necessary for transcriptional activity of HIF-1 complex.

Requires phosphorylation for DNA-binding. Phosphorylation at Ser-247 by CSNK1D/CK1 represses kinase activity and impairs ARNT binding. Phosphorylation by GSK3-beta and PLK3 promote degradation by the proteasome.

Sumoylated; with SUMO1 under hypoxia. Sumoylation is enhanced through interaction with RWDD3. Both sumoylation and desumoylation seem to be involved in the regulation of its stability during hypoxia. Sumoylation can promote either its stabilization or its VHL-dependent degradation by promoting hydroxyproline-independent HIF1A-VHL complex binding, thus leading to HIF1A ubiquitination and proteasomal degradation. Desumoylation by SENP1 increases its stability amd transcriptional activity. There is a disaccord between various publications on the effect of sumoylation and desumoylation on its stability and transcriptional activity.

Acetylation of Lys-532 by ARD1 increases interaction with VHL and stimulates subsequent proteasomal degradation (PubMed:12464182). Deacetylation of Lys-709 by SIRT2 increases its interaction with and hydroxylation by EGLN1 thereby inactivating HIF1A activity by inducing its proteasomal degradation (PubMed:24681946).

Polyubiquitinated; in normoxia, following hydroxylation and interaction with VHL. Lys-532 appears to be the principal site of ubiquitination. Clioquinol, the Cu/Zn-chelator, inhibits ubiquitination through preventing hydroxylation at Asn-803. Ubiquitinated by a CUL2-based E3 ligase.

The iron and 2-oxoglutarate dependent 3-hydroxylation of asparagine is (S) stereospecific within HIF CTAD domains. [UniProt]

Images (3) Click the Picture to Zoom In

  • ARG41701 anti-HIF-1 alpha antibody ICC/IF image

    Immunofluorescence: HeLa cells treated with chloride. Cells were stained with ARG41701 anti-HIF-1 alpha antibody.

  • ARG41701 anti-HIF-1 alpha antibody IHC-P image

    Immunohistochemistry: Paraffin-embedded Human kindey tissue stained with ARG41701 anti-HIF-1 alpha antibody.

  • ARG41701 anti-HIF-1 alpha antibody WB image

    Western blot: Ramos cell lysate stained with ARG41701 anti-HIF-1 alpha antibody.