Product Pathways - Protein Translation
HIF-1α (D2U3T) Rabbit mAb #14179
|14179S||100 µl ( 10 western blots )||￥3,250.00 现货查询||购买询价|
|14179||carrier free & custom formulation / quantity||email request|
Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting, ChIP=Chromatin IP, ChIP-seq=Chromatin IP-seq,
Specificity / Sensitivity
HIF-1α (D2U3T) Rabbit mAb recognizes endogenous levels of total HIF-1α protein.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Lys460 of human HIF-1α protein.
Western blot analysis of extracts from various cell lines, untreated (-) or treated with either DMOG (1 mM, 6 hr; +) or cobalt chloride (100 nM, 4 hr; +) as indicated, using HIF-1α (D2U3T) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Chromatin immunopreciptations were performed with cross-linked chromatin from 4 x 106 MCF7 cells treated with cobalt chloride (100 μM) overnight and either 10 μl of HIF-1α (D2U3T) Rabbit mAb or 2 μL of Normal Rabbit IgG #2729, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human ARRDC3 Downstream Primers #75671, human ERRFI1 upstream primers, and SimpleChIP® Human α Satelite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 MCF7 cells treated with cobalt chloride (100 μM) overnight and 10 μl of HIF-1α (D2U3T) Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared from 5 ng enriched ChIP DNA using NEBNext® Ultra™ II DNA Library Prep Kit for Illumina®, and sequenced on the Illumina NextSeq. The figure shows binding across the PCAT6 gene. For additional ChIP-seq tracks, please download the product data sheet.
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 (VLH) 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).
- Sharp, F.R. and Bernaudin, M. (2004) Nat Rev Neurosci 5, 437-48.
- Wang, G.L. et al. (1995) Proc Natl Acad Sci U S A 92, 5510-4.
- Jaakkola, P. et al. (2001) Science 292, 468-72.
- Maxwell, P.H. et al. (1999) Nature 399, 271-5.
- Fukuda, R. et al. (2002) J Biol Chem 277, 38205-11.
- Jiang, B.H. et al. (2001) Cell Growth Differ 12, 363-9.
- Laughner, E. et al. (2001) Mol Cell Biol 21, 3995-4004.
- Walisser, J.A. et al. (2004) Proc Natl Acad Sci U S A 101, 16677-82.
- Salomon-Nguyen, F. et al. (2000) Proc Natl Acad Sci U S A 97, 6757-62.
- Gunton, J.E. et al. (2005) Cell 122, 337-49.
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