Revision 1
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.
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UniProt ID:

#Q16539, #O15264, #P53778, #Q15759

Entrez-Gene Id:

1432, 5603, 6300, 5600

Product Information

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 MAPK Antibody detects phosphorylated threonine 180 and tyrosine 182 of p38 MAPK but does not appreciably cross-react with the corresponding phosphorylated forms of either p42/44 MAPK or SAPK/JNK. p38 MAPK (D13E1) XP® Rabbit mAb recognizes endogenous levels of total p38α, ß, or γ MAPK protein and does not recognize p38δ, SAPK/JNK, or p44/42 MAPK proteins.

Source / Purification

Monoclonal activation state antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr180/Tyr182 of human p38 MAPK. Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human p38 protein.

Product Description

The PhosphoPlus® p38 MAPK (Thr180/Tyr182) Antibody Kit provides reagents and protocols for the rapid analysis of the phosphorylation status of p38 MAPK at Thr180/Tyr182.

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).

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.
PhosphoPlus is a registered 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

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Revision 1
#9210

PhosphoPlus® p38 MAPK (Thr180/Tyr182) Antibody Kit

PhosphoPlus® p38 MAPK (Thr180/Tyr182) Antibody Kit: Image 1 Expand Image
使用 Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb(上)或 p38 MAPK Antibody # 9212(下),对未经处理或经紫外线处理的 COS 和 293 细胞提取物进行蛋白质印迹分析。
PhosphoPlus® p38 MAPK (Thr180/Tyr182) Antibody Kit: Image 2 Expand Image
一抗与靶标蛋白结合之后,与偶联 HRP 的二抗形成复合体。添加 LumiGLO®,在酶催化分解期间发光。
PhosphoPlus® p38 MAPK (Thr180/Tyr182) Antibody Kit: Image 3 Expand Image
使用 p38 MAPK (D13E1) XP® Rabbit mAb 对不同细胞系提取物进行蛋白质印迹分析。
PhosphoPlus® p38 MAPK (Thr180/Tyr182) Antibody Kit: Image 4 Expand Image
使用 Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb 对 p38 MAPK Control Cell extracts 进行蛋白质印迹分析。

对p38 MAPK 对照细胞提取物进行蛋白质印迹法分析。

PhosphoPlus® p38 MAPK (Thr180/Tyr182) Antibody Kit: Image 5 Expand Image
使用 Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb 对石蜡包埋的人结肠癌进行免疫组织化学分析。
PhosphoPlus® p38 MAPK (Thr180/Tyr182) Antibody Kit: Image 6 Expand Image
使用 p38 MAPK (D13E1) XP® Rabbit mAb 对石蜡包埋的人乳腺癌细胞进行免疫组织化学分析。
PhosphoPlus® p38 MAPK (Thr180/Tyr182) Antibody Kit: Image 7 Expand Image
使用 Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb 对石蜡包埋的小鼠结肠进行免疫组织化学分析。
PhosphoPlus® p38 MAPK (Thr180/Tyr182) Antibody Kit: Image 8 Expand Image
使用 p38 MAPK (D13E1) XP® Rabbit mAb 对石蜡包埋的人结肠癌细胞进行免疫组织化学分析。
PhosphoPlus® p38 MAPK (Thr180/Tyr182) Antibody Kit: Image 9 Expand Image
使用 Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb,对未经处理(左)或经紫外线处理(右)的石蜡包埋的 293T 细胞沉淀物进行免疫组织化学分析。
PhosphoPlus® p38 MAPK (Thr180/Tyr182) Antibody Kit: Image 10 Expand Image
在对照肽(左)或抗原特异性肽(右)存在的情况下,使用 p38 MAPK (D13E1) XP® Rabbit mAb 对石蜡包埋的人肺癌细胞进行免疫组织化学分析。
PhosphoPlus® p38 MAPK (Thr180/Tyr182) Antibody Kit: Image 11 Expand Image
使用 Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb(绿色),对未经处理(左)或经 anisomycin 处理(右)的 COS 细胞进行共聚焦免疫荧光分析。肌动蛋白纤丝用 DY-554 Phalloidin 进行标记(红色)。
PhosphoPlus® p38 MAPK (Thr180/Tyr182) Antibody Kit: Image 12 Expand Image
使用 p38 MAPK (D13E1) XP® Rabbit mAb(绿色)对未经(左)或经紫外线处理(100 mJ/cm2,30 分钟恢复;右)的 HeLa 细胞进行共聚焦免疫荧光分析。使用 DY-554 phalloidin(红色)标记肌动蛋白纤丝。
PhosphoPlus® p38 MAPK (Thr180/Tyr182) Antibody Kit: Image 13 Expand Image
使用 Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb(实线)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线),对未经处理(蓝色)或经 Anisomycin(25 µM,30 分钟;绿色)处理的 Jurkat 细胞进行流式细胞分析。Anti-rabbit IgG (H+L)、F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 作为二抗。
PhosphoPlus® p38 MAPK (Thr180/Tyr182) Antibody Kit: Image 14 Expand Image
以浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)作为对照,使用 p38 MAPK (D13E1) XP® Rabbit mAb(实线)对 Hela 细胞进行流式细胞分析。Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 用作二抗。