ARG10515

anti-HIF-1 alpha antibody [ESEE122]

anti-HIF-1 alpha antibody [ESEE122] for ELISA,Flow cytometry,ICC/IF,IHC-Formalin-fixed paraffin-embedded sections,IHC-Frozen sections,Western blot and Bovine,Human,Mouse,Rat

Cancer antibody; Cell Biology and Cellular Response antibody; Gene Regulation antibody; Metabolism antibody
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Overview

Product Description

Mouse Monoclonal antibody [ESEE122] recognizes HIF-1 alpha

Tested Reactivity Hu, Ms, Rat, Bov
Tested Application ELISA, FACS, ICC/IF, IHC-Fr, IHC-P, WB
Host Mouse
Clonality Monoclonal
Clone ESEE122
Isotype IgG1
Target Name HIF-1 alpha
Antigen Species Human
Immunogen Synthetic peptide within 329 to 530 aa of Human HIF-1A protein (NP_001230013.1)
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
ELISAAssay-dependent.
FACS1µg for 10^6  cells.
ICC/IF5 - 12 µg/ml
IHC-Fr1:1000 - 1:8000
IHC-P1:1000 - 1:8000
WB1:100 - 1:1000
Application Note IHC-P: Antigen Retrieval: By heat mediation.
* The dilutions indicate recommended starting dilutions and the optimal dilutions or concentrations should be determined by the scientist.
Positive Control Hypoxic cell pellets.

Properties

Form Liquid
Purification Purified by affinity chromatography.
Buffer PBS and 0.02% Sodium azide
Preservative 0.02% Sodium azide
Concentration 1 mg/ml
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 or below. 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: 15251 Mouse HIF1A

GeneID: 29560 Rat HIF1A

GeneID: 3091 Human HIF1A

Gene Symbol HIF1A
Gene Full Name hypoxia inducible factor 1, alpha subunit (basic helix-loop-helix transcription factor)
Background 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]
Highlight Related products:
HIF1 alpha antibodies; HIF1 alpha ELISA Kits; Anti-Mouse IgG secondary antibodies;
Related news:
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Hypoxia-induced transcription, histone demethylases are involved
Research Area Cancer antibody; Cell Biology and Cellular Response antibody; Gene Regulation antibody; Metabolism antibody
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.

Customer's Feedback

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Review for anti-HIF-1 alpha antibody [ESEE122]

Application:IHC

Sample:Mouse muscle tissue

Sample Preparation Method:Paraffin-embedded

Primary Antibody Dilution Factor:1:90

Primary Antibody Incubation Time:2 hours

Primary Antibody Incubation Temperature:37 ºC

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Review for anti-HIF-1 alpha antibody [ESEE122]

Application:IF/ICC

Sample:NRK-49F untreated or treated with CoCl2 (400 μM) for 8 h

Fixation Buffer:4% PFA

Fixation Time:15 min

Fixation Temperature:20 - 25 ºC

Permeabilization Buffer:0.5% Triton X-100

Primary Antibody Dilution Factor:1:200

Primary Antibody Incubation Time:overnight

Primary Antibody Incubation Temperature:4 ºC

Conjugation of Secondary Antibody:Alexa Fluor 594

Specific References

AKT, a Key Transmitter of HIF-1α and AR Signaling Pathways, Has a Critical Role in the Apigetrin-Mediated Anti-Cancer Effects in Prostate Cancer Cells

WB / Human

You-Kyung Lee et al.
Biomedicines,  (2022)

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Huperzine A, reduces brain iron overload and alleviates cognitive deficit in mice exposed to chronic intermittent hypoxia.

WB / Mouse

An Ji-Ren et al.
Life Sci.,  (2020)

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