| Product Includes | Product # | Quantity | Color | Storage Temp |
|---|---|---|---|---|
| HIF-1α Goat Antibody Coated Microwells | 25917 | 96 tests |
|
+4C |
| HIF-1α Rabbit Detection mAb | 14714 | 1 ea |
|
+4C |
| Anti-rabbit IgG, HRP-linked Antibody (ELISA Formulated) | 13272 | 1 ea |
|
+4C |
| Detection Antibody Diluent | 13339 | 11 ml |
|
+4C |
| HRP Diluent | 13515 | 11 ml |
|
+4C |
| TMB Substrate | 7004 | 11 ml |
|
+4C |
| STOP Solution | 7002 | 11 ml |
|
+4C |
| Sealing Tape | 54503 | 2 ea |
|
+4C |
| ELISA Wash Buffer (20X) | 9801 | 25 ml |
|
+4C |
| ELISA Sample Diluent | 11083 | 25 ml |
|
+4C |
| Cell Lysis Buffer (10X) | 9803 | 15 ml |
|
-20C |
*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.
Description
The PathScan® Total HIF-1α Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total HIF-1α protein. A HIF-1α antibody has been coated onto the microwells. After incubation with cell lysates, HIF-1α protein is captured by the coated antibody. Following extensive washing, an HIF-1α Detection Antibody is added to detect the captured HIF-1α protein. Anti-Rabbit IgG, HRP-linked Antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of optical density for this developed color is proportional to the quantity of HIF-1α protein.
*Antibodies in this kit are custom formulations specific to kit.
Specificity/Sensitivity
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).
- 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.
Background References
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
Trademarks and Patents
Limited Uses
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