Cell Signaling Technology

Product Pathways - Protein Translation

HIF-1α (D5F3M) Mouse mAb #79233

ARNT interacting protein   ARNT-interacting protein   Basic-helix-loop-helix-PAS protein MOP1   BHLHE78   Class E basic helix-loop-helix protein 78   HIF-1-alpha   Hif-1a   HIF-1alpha   HIF1   HIF1-alpha   HIF1A   hypoxia inducible factor 1. alpha subunit (basic helix-loop-helix transcription factor)   hypoxia-inducible factor 1 alpha isoform I.3   Hypoxia-inducible factor 1-alpha   hypoxia-inducible factor 1. alpha subunit (basic helix-loop-helix transcription factor)   Member of PAS protein 1   member of PAS superfamily 1   MOP1   PAS domain-containing protein 8   PASD8  

No. Size Price
79233S 100 µl ( 10 western blots ) ¥3,250.00 现货查询 购买询价
79233T 20 µl ( 2 western blots ) ¥1,200.00 现货查询 购买询价
79233 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Monkey, Endogenous 120 Mouse IgG1
IP 1:50
F 1:1600
IF-IC 1:800

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, F=Flow Cytometry, IF-IC=Immunofluorescence (Immunocytochemistry),

Specificity / Sensitivity

HIF-1α (D5F3M) Mouse mAb recognizes endogenous levels of total HIF-1α protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the carboxy terminus of human HIF-1α protein.

IF-IC

IF-IC

Confocal immunofluorescent analysis of Hep G2 cells, untreated (left) or treated with cobalt chloride (500 μM, 24 h; right), using HIF-1α (D5F3M) Mouse mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of U-2 OS cells, untreated (blue) or treated with DMOG (1mM, 6 h; green) using HIF-1α (D5F3M) Mouse mAb. Anti-mouse IgG (H+L), F(ab')2 fragment (Alexa Fluor® 488 conjugate) #4408 was used as a secondary antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from Hep G2 cells untreated or treated with cobalt chloride (100 µM, 4 h; +) and U-2 OS cells untreated or treated with DMOG (1 mM, 6 h; +) using HIF-1α Mouse (D5F3M) Mouse mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

Background

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 (VHL) 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).

  1. Sharp, F.R. and Bernaudin, M. (2004) Nat Rev Neurosci 5, 437-48.
  2. Wang, G.L. et al. (1995) Proc Natl Acad Sci U S A 92, 5510-4.
  3. Jaakkola, P. et al. (2001) Science 292, 468-72.
  4. Maxwell, P.H. et al. (1999) Nature 399, 271-5.
  5. Fukuda, R. et al. (2002) J Biol Chem 277, 38205-11.
  6. Jiang, B.H. et al. (2001) Cell Growth Differ 12, 363-9.
  7. Laughner, E. et al. (2001) Mol Cell Biol 21, 3995-4004.
  8. Walisser, J.A. et al. (2004) Proc Natl Acad Sci U S A 101, 16677-82.
  9. Salomon-Nguyen, F. et al. (2000) Proc Natl Acad Sci U S A 97, 6757-62.
  10. Gunton, J.E. et al. (2005) Cell 122, 337-49.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

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Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Alexa Fluor is a registered trademark of Life Technologies Corporation.

Tween is a registered trademark of ICI Americas, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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