Phospho-p38 MAPK (Thr180/Tyr182) (28B10) Mouse mAb (Alexa Fluor® 647 Conjugate) #4552
Filter:
- F
Supporting Data
| REACTIVITY | H M R Mk Sc |
| SENSITIVITY | Endogenous |
| MW (kDa) | |
| Source/Isotype | Mouse IgG1 |
Application Key:
- F-Flow Cytometry
Species Cross-Reactivity Key:
- H-Human
- M-Mouse
- R-Rat
- Mk-Monkey
- Sc-S. cerevisiae
Product Information
Product Description
This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 647 fluorescent dye and tested in-house for direct flow cytometric analysis of human and mouse cells. The unconjugated antibody #9216 reacts with human, mouse, rat, D. melanogaster, monkey, S.cerevisiae and zebra fish phospho-p38 MAPK (Thr180/Tyr182). CST expects that Phospho-p38 MAPK (Thr180/Tyr182) (28B10) Mouse mAb (Alexa Fluor® 647 Conjugate) will also recognize phospho-p38 MAPK (Thr180/Tyr182) in these species.
Product Usage Information
| Application | Dilution |
|---|---|
| Flow Cytometry (Fixed/Permeabilized) | 1:50 |
Storage
Supplied in PBS (pH 7.2), less than 0.1% sodium azide and 2 mg/ml BSA. Store at 4°C. Do not aliquot the antibody. Protect from light. Do not freeze.
Protocol
Specificity / Sensitivity
Phospho-p38 MAPK (Thr180/Tyr182) (28B10) Mouse mAb (Alexa Fluor® 647 Conjugate) detects endogenous levels of p38 MAP kinase only when dually phosphorylated at Thr180 and Tyr182. This antibody does not appreciably cross-react with the corresponding phosphorylated forms of either p44/42 MAPK (Erk1/2) or SAPK/JNK.
Species Reactivity:
Human, Mouse, Rat, Monkey, S. cerevisiae
The antigen sequence used to produce this antibody shares 100% sequence homology with the species listed here, but reactivity has not been tested or confirmed to work by CST. Use of this product with these species is not covered under our Product Performance Guarantee.
Species predicted to react based on 100% sequence homology:
Zebrafish
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr180/Tyr182 of human p38 MAP kinase. This antibody was conjugated to Alexa Fluor® 647 under optimal conditions with an F/P ratio of 2-6. The Alexa Fluor® 647 dye is maximally excited by red light (e.g. 633 nm He-Ne laser). Antibody conjugates of the Alexa Fluor® 647 dye produce bright far-red-fluorescence emission, with a peak at 665 nm.
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.
限制使用
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For Research Use Only. Not For Use In Diagnostic Procedures.
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