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48-602MAG
Buffer Detection Kit for Magnetic Beads
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Anti-Hypoxia Inducible Factor 1 α Antibody, clone H1α67 is a Mouse Monoclonal Antibody for detection of Hypoxia Inducible Factor 1 α also known as HIF-1 alpha or ARNT Interacting Protein & has been validated in EMSA, IHC, IP & WB.
More>>Anti-Hypoxia Inducible Factor 1 α Antibody, clone H1α67 is a Mouse Monoclonal Antibody for detection of Hypoxia Inducible Factor 1 α also known as HIF-1 alpha or ARNT Interacting Protein & has been validated in EMSA, IHC, IP & WB. Less<<
HIF is a transcriptional complex that plays a central role in mammalian oxygen homeostasis, the posttranslational modification by prolyl hydroxylation as a key regulatory event that targets HIF-alpha subunits for proteasomal destruction via the von Hippel-Lindau ubiquitylation complex. The transcriptional complex is composed of an alpha-beta heterodimer; HIF-beta being a constitutive nuclear protein that dimerises with oxygen regulated HIF-alpha subunits. In normoxia, 4-hydroxylation of human HIF-alpha at Pro402 or Pro564 by a set of HIF prolyl hydroxylase isoenzymes (PHD 1-3) mediates HIF1-alpha recognition by von Hippel-Lindau ubiquitin ligase complex leading to its proteasomal destruction. In hypoxia (deprivation of oxygen), lack of hydroxylase activity enables HIF-alpha subunits to escape destruction and become transcriptionally active. Thus HIF hydroxylases provide a focus for understanding cellular responses to hypoxia and target for therapeutic manipulation. There are several HIF factors, which include HIF 1-alpha, HIF 1-beta, HIF 2-alpha. HIF 1-alpha is an 812 a.a. protein in rat and 836 a.a. long in mouse and human. 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, 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. It is ubiquitous in expression as cytoplasmic in normoxia, nuclear translocation in response to hypoxia.
References
Product Information
Format
Purified
Control
Cobalt chloride-treated MCF-7 cells
Presentation
Protein A Purified mouse immunoglobulin in 20 mM sodium phosphate, 250 mM NaCl, pH. 7.6, with 0.1% sodium azide as a preservative.
Anti-Hypoxia Inducible Factor 1 α Antibody, clone H1α67 is a Mouse Monoclonal Antibody for detection of Hypoxia Inducible Factor 1 α also known as HIF-1 alpha or ARNT Interacting Protein & has been validated in EMSA, IHC, IP & WB.
Key Applications
Electrophoretic Mobility Shift Assay
Immunohistochemistry
Immunoprecipitation
Western Blotting
Application Notes
Western blot: 1:500-1:1,000. The antibody recognizes a band of 120 kD in induced tissues and cells. Multiple bands may be present at 120 kD representing post-translational modification of HIF-1alpha.
Immunohistochemistry: 1:500-1:1,000. The antibody has been used successfully on formalin-fixed, paraffin embedded tissue sections after antigen retrieval.
Immunoprecipitation
Gel Shift
Optimal working dilutions must be determined by end user.
Biological Information
Immunogen
Fusion protein from amino acids 432-528 of human HIF-1alpha.
Hypoxia-inducible factor-1 (HIF1) is a transcription factor found in mammalian cells cultured under reduced oxygen tension that plays an essential role in cellular and systemic homeostatic responses to hypoxia. HIF1 is a heterodimer composed of an alpha subunit and a beta subunit. The beta subunit has been identified as the aryl hydrocarbon receptor nuclear translocator (ARNT). This gene encodes the alpha subunit of HIF-1. Overexpression of a natural antisense transcript (aHIF) of this gene has been shown to be associated with nonpapillary renal carcinomas. Two alternative transcripts encoding different isoforms have been identified.
FUNCTION: SwissProt: Q16665 # 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, 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. SIZE: 826 amino acids; 92670 Da SUBUNIT: Interacts with COPS5 subunit of COP9 signalosome complex, leading to the regulation of its stability. Interacts with TSGA10 (By similarity). Efficient DNA binding requires heterodimerization of an alpha and a beta/ARNT subunit. Binds to the TAZ-type 1 domains of CREBBP and EP300. Interacts with NCOA1, NCOA2, APEX and HSP90. Interacts with VHL which docks HFA1 to the E3 ubiquitin ligase complex for subsequent destruction. Interaction, via the ODD domain, with the beta domain of VHLL, protects HIF1A from destruction by competing against the destructive targeting initiated by VHL. SUBCELLULAR LOCATION: Cytoplasm. Nucleus. Note=Cytoplasmic in normoxia, nuclear translocation in response to hypoxia. TISSUE SPECIFICITY: Expressed in most tissues with highest levels in kidney and heart. Overexpressed in the majority of common human cancers and their metastases, due to the presence of intratumoral hypoxia and as a result of mutations in genes encoding oncoproteins and tumor suppressors.DOMAIN:SwissProt: Q16665 Contains two independent C-terminal transactivation domains, NTAD and CTAD, which function synergistically. Their transcriptional activity is repressed by an intervening inhibitory domain (ID). PTM: In normoxia, is hydroxylated on Pro-402 and Pro-564 in the oxygen-dependent degradation domain (ODD) by EGLN1/PHD1 and EGLN2/PHD2. EGLN3/PHD3 has also been shown to hydroxylate Pro-564. The hydroxylated prolines promote interaction with VHL, initiating rapid ubiquitination and subsequent proteasomal degradation. 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. & S-nitrosylation of Cys-800 may be responsible for increased recruitment of p300 coactivator necessary for transcriptional activity of HIF-1 complex. & Acetylation of Lys-532 by ARD1 increases interaction with VHL and stimulates subsequent proteasomal degradation. & Requires phosphorylation for DNA-binding. SIMILARITY: Contains 1 basic helix-loop-helix (bHLH) domain. & Contains 1 PAC (PAS-associated C-terminal) domain. & Contains 2 PAS (PER-ARNT-SIM) domains.
Molecular Weight
Predicted MW: 96 kDa, apparent MW: 120 kDa
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality Assurance
Tested
Usage Statement
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Storage and Shipping Information
Storage Conditions
Maintain for 1 year at 2–8°C from date of shipment. Aliquot to avoid repeated freezing and thawing. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
