Revision 3
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IHC-P, IF-IC

REACTIVITY:

H M R Mk Dm

SENSITIVITY:

Endogenous

MW (kDa):

43

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q16539, #O15264, #P53778, #Q15759

Entrez-Gene Id:

1432, 5603, 6300, 5600

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunohistochemistry (Paraffin) 1:100
Immunofluorescence (Immunocytochemistry) 1:100

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

Phospho-p38 MAP Kinase (Thr180/Tyr182) (12F8) Rabbit mAb detects endogenous levels of p38 MAPK only when dually phosphorylated at threonine 180 and tyrosine182. It will also react with p38 singly phosphorlyated at Thr180. This antibody does not cross-react with the phosphorylated forms of either p42/44 MAPK or SAPK/JNK.

Species Reactivity:

Human, Mouse, Rat, Monkey, D. melanogaster

Species predicted to react based on 100% sequence homology

Hamster, Mink, Zebrafish, Horse

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr180/Tyr182 of human p38 MAPK.

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).

  1. Rouse, J. et al. (1994) Cell 78, 1027-37.
  2. Han, J. et al. (1994) Science 265, 808-11.
  3. Lee, J.C. et al. (1994) Nature 372, 739-46.
  4. Freshney, N.W. et al. (1994) Cell 78, 1039-49.
  5. Raingeaud, J. et al. (1995) J Biol Chem 270, 7420-6.
  6. Zervos, A.S. et al. (1995) Proc Natl Acad Sci U S A 92, 10531-4.
  7. Zhao, M. et al. (1999) Mol Cell Biol 19, 21-30.
  8. Yang, S.H. et al. (1999) Mol Cell Biol 19, 4028-38.
  9. Cuenda, A. et al. (1995) FEBS Lett 364, 229-33.
  10. Kumar, S. et al. (1999) Biochem Biophys Res Commun 263, 825-31.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry)

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

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Revision 3
#4631

Phospho-p38 MAPK (Thr180/Tyr182) (12F8) Rabbit mAb

Western Blotting Image 1: Phospho-p38 MAPK (Thr180/Tyr182) (12F8) Rabbit mAb Expand Image
使用 Phospho-p38 MAPK (Thr180/Tyr182) (12F8) Rabbit mAb,对未经处理或按指定要求处理的 Jurkat、NIH/3T3 和 pC12 细胞提取物进行蛋白质印迹分析。
Immunohistochemistry Image 1: Phospho-p38 MAPK (Thr180/Tyr182) (12F8) Rabbit mAb Expand Image
使用 Phospho-p38 MAPK (Thr180/Tyr182) (12F8) Rabbit mAb 对石蜡包埋肺癌进行免疫组织化学分析,显示核定位。
Immunohistochemistry Image 2: Phospho-p38 MAPK (Thr180/Tyr182) (12F8) Rabbit mAb Expand Image
使用 Phospho-p38 MAPK (Thr180/Tyr182) (12F8) Rabbit mAb 对石蜡包埋结肠癌进行免疫组织化学分析,显示核定位。
Immunohistochemistry Image 3: Phospho-p38 MAPK (Thr180/Tyr182) (12F8) Rabbit mAb Expand Image
使用 Phospho-p38 MAPK (Thr180/Tyr182) (12F8) Rabbit mAb,对未经处理 (左图) 或 CIP 磷酸酶处理 (右图) 的石蜡包埋人胶质母细胞瘤进行免疫组织化学分析。

Immunohistochemistry Image 4: Phospho-p38 MAPK (Thr180/Tyr182) (12F8) Rabbit mAb Expand Image
使用 Phospho-p38 MAPK (Thr180/Tyr182) (12F8) Rabbit mAb (左图) 或使用抗原磷酸肽预孵育的相同抗体 (右图),对石蜡包埋的乳腺癌进行免疫组织化学分析。
Immunohistochemistry Image 5: Phospho-p38 MAPK (Thr180/Tyr182) (12F8) Rabbit mAb Expand Image
使用 Phospho-p38 MAPK (Thr180/Tyr182) (12F8) Rabbit mAb,对未经处理 (左图) 或 Anisomycin (右图) 的石蜡包埋 NIH/3T3 细胞进行免疫组织化学分析。
Immunofluorescence Image 1: Phospho-p38 MAPK (Thr180/Tyr182) (12F8) Rabbit mAb Expand Image
使用 Phospho-p38 MAPK (绿色) 对-/+λ 紫外线处理的 HeLa 细胞进行共聚焦免疫荧光分析。未经处理细胞 (左图) 无染色及经处理细胞在核定位 (右图) 。红色 = 肌动蛋白丝 (phalloidin)。