General description
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.
Specificity
Recognizes Hypoxia inducible factor-1alpha (HIF-1alpha).
Immunogen
Fusion protein from amino acids 432-528 of human HIF-1alpha.
Application
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.
Research Category
Epigenetics & Nuclear Function
Research Sub Category
Transcription Factors
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.
Target description
Predicted MW: 96 kDa, apparent MW: 120 kDa
Physical form
Format: Purified
Protein A Purified mouse immunoglobulin in 20 mM sodium phosphate, 250 mM NaCl, pH. 7.6, with 0.1% sodium azide as a preservative.
Protein A purified
Storage and Stability
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.
Analysis Note
Control
Cobalt chloride-treated MCF-7 cells
Other Notes
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Legal Information
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
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.