| Product Includes | Product # | Quantity | Color |
|---|---|---|---|
| FastScan™ ELISA Microwell Strip Plate, 96 Well | 53257 | 96 tests | |
| p38α MAPK Rabbit Capture mAb | 31880 | 1 ea | Green (Lyophilized) |
| p38α MAPK Mouse HRP-linked mAb | 28227 | 1 ea | Red (Lyophilized) |
| FastScan™ ELISA Capture Antibody Diluent | 16076 | 3 ml | Green |
| FastScan™ ELISA HRP Antibody Diluent | 28120 | 3 ml | |
| TMB Substrate | 7004 | 11 ml | |
| STOP Solution | 7002 | 11 ml | |
| Sealing Tape | 54503 | 1 ea | |
| ELISA Wash Buffer (20X) | 9801 | 25 ml | |
| FastScan™ ELISA Cell Extraction Buffer (5X) | 69905 | 10 ml | |
| FastScan™ ELISA Cell Extraction Enhancer Solution (50X) | 25243 | 1 ml | |
| FastScan™ ELISA Kit #15401 Positive Control Type 2 | 33430 | 1 ea |
*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.
Description
The FastScan™ Total p38α MAPK ELISA Kit is a sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of p38α MAPK protein. To perform the assay, sample is incubated with a capture antibody conjugated with a proprietary tag and a second detection antibody linked to HRP, forming a sandwich with p38α MAPK in solution. This entire complex is immobilized to the plate via an anti-tag antibody. The wells are then washed to remove unbound material. TMB is then added. The magnitude of observed signal is proportional to the quantity of p38α MAPK protein.
*Antibodies in this kit are custom formulations specific to kit.
IMPORTANT: This FastScan™ ELISA Kit requires 4 washes at Step 6 of the protocol.
Specificity/Sensitivity
Background
p38 MAP kinase (MAPK), also called RK (1) or CSBP (2), is the mammalian orthologue of the yeast HOG kinase that participates in a signaling cascade controlling cellular responses to cytokines and stress (1-4). Four isoforms of p38 MAPK, p38α, β, γ (also known as Erk6 or SAPK3), and δ (also known as SAPK4) have been identified. Similar to the SAPK/JNK pathway, p38 MAPK is activated by a variety of cellular stresses, including osmotic shock, inflammatory cytokines, lipopolysaccharide (LPS), UV light, and growth factors (1-5). MKK3, MKK6, and SEK activate p38 MAPK by phosphorylation at Thr180 and Tyr182. Activated p38 MAPK has been shown to phosphorylate and activate MAPKAP kinase 2 (3) and to phosphorylate the transcription factors ATF-2 (5), Max (6), and MEF2 (5-8). SB203580 (4-(4-fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)-imidazole) is a selective inhibitor of p38 MAPK. This compound inhibits the activation of MAPKAPK-2 by p38 MAPK and subsequent phosphorylation of HSP27 (9). SB203580 inhibits p38 MAPK catalytic activity by binding to the ATP-binding pocket, but does not inhibit phosphorylation of p38 MAPK by upstream kinases (10).
- Rouse, J. et al. (1994) Cell 78, 1027-37.
- Han, J. et al. (1994) Science 265, 808-11.
- Lee, J.C. et al. (1994) Nature 372, 739-46.
- Freshney, N.W. et al. (1994) Cell 78, 1039-49.
- Raingeaud, J. et al. (1995) J Biol Chem 270, 7420-6.
- Zervos, A.S. et al. (1995) Proc Natl Acad Sci U S A 92, 10531-4.
- Zhao, M. et al. (1999) Mol Cell Biol 19, 21-30.
- Yang, S.H. et al. (1999) Mol Cell Biol 19, 4028-38.
- Cuenda, A. et al. (1995) FEBS Lett 364, 229-33.
- Kumar, S. et al. (1999) Biochem Biophys Res Commun 263, 825-31.
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
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