Revision 1

#9790Store at -20C

1 Kit

(5 x 20 microliters)

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.
Product Includes Product # Quantity Mol. Wt Isotype/Source
p38α MAPK Antibody 9218 20 µl 40 kDa Rabbit 
p38β MAPK (C28C2) Rabbit mAb 2339 20 µl 43 kDa Rabbit IgG
p38γ MAPK Antibody 2307 20 µl 46 kDa Rabbit 
p38δ MAPK (10A8) Rabbit mAb 2308 20 µl 43 kDa Rabbit IgG
Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb 4511 20 µl 43 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The p38 MAPK Isoform Activation Antibody Sampler Kit provides an economical means to evaluate the activation status of individual isoforms of p38 MAPK through immunoprecipitation of the phosphorylated p38 MAPK followed by western blot using isoform specific antibodies. The kit includes enough primary and secondary antibodies to perform two IP/western blot experiments.

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.

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.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    Limited Uses

    Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

    Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

    Revision 1
    #9790

    p38 MAPK Isoform Activation Antibody Sampler Kit

    p38 MAPK Isoform Activation Antibody Sampler Kit: Image 1 Expand Image
    Western blot analysis of purified recombinant full-length p38 MAPK GST fusion protein, using p38 MAP kinase pan antibody (upper) or p38γ MAPK Antibody (lower).
    p38 MAPK Isoform Activation Antibody Sampler Kit: Image 2 Expand Image
    Western blot analysis of purified recombinant full-length p38 MAPK GST fusion proteins, using p38 MAPK pan antibody (upper), or p38δ MAPK (10A8) Rabbit mAb (lower).
    p38 MAPK Isoform Activation Antibody Sampler Kit: Image 3 Expand Image
    Western blot analysis of purified recombinant full-length p38 MAPK GST-fusion proteins, using p38β MAPK (C28C2) Rabbit mAb (upper), or a p38 MAPK pan antibody (lower).
    p38 MAPK Isoform Activation Antibody Sampler Kit: Image 4 Expand Image
    Western blot analysis of extracts from COS and 293 cells, untreated or UV-treated, using Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb (upper) or p38 MAPK Antibody #9212 (lower).
    p38 MAPK Isoform Activation Antibody Sampler Kit: Image 5 Expand Image
    After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
    p38 MAPK Isoform Activation Antibody Sampler Kit: Image 6 Expand Image
    Western blot analysis of extracts from HeLa, NIH/3T3, PC12 and COS cells, using p38α MAPK Antibody.
    p38 MAPK Isoform Activation Antibody Sampler Kit: Image 7 Expand Image
    Western blot analysis of extracts from HUVEC and U937 cells, using p38γ MAPK Antibody.
    p38 MAPK Isoform Activation Antibody Sampler Kit: Image 8 Expand Image
    Western blot analysis of extracts from 293, NBT-II and PC12 cells, using p38δ MAPK (10A8) Rabbit mAb.
    p38 MAPK Isoform Activation Antibody Sampler Kit: Image 9 Expand Image
    Westen blot analysis of extracts from HUVEC and COS cells using p38β MAPK (C28C2) Rabbit mAb.
    p38 MAPK Isoform Activation Antibody Sampler Kit: Image 10 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb.
    p38 MAPK Isoform Activation Antibody Sampler Kit: Image 11 Expand Image
    Western blot analysis of extracts from Xenopus oocytes overexpressing Myc-tagged p38 isoforms, using p38α MAPK Antibody (upper) and Myc-Tag Antibody (lower) to demonstrate the isoform specificity of p38α MAPK Antibody. (Provided by Dr. Eusebio Perdiguero and Dr. Angel Nebreda, European Molecular Biology Laboratory, Germany.)
    p38 MAPK Isoform Activation Antibody Sampler Kit: Image 12 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse colon using Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb.
    p38 MAPK Isoform Activation Antibody Sampler Kit: Image 13 Expand Image
    Immunohistochemical analysis of paraffin-embedded 293T cell pellets, untreated (left) or UV-treated (right), using Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb.
    p38 MAPK Isoform Activation Antibody Sampler Kit: Image 14 Expand Image
    Confocal immunofluorescent analysis of COS cells, untreated (left) or anisomycin-treated (right) using Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).
    p38 MAPK Isoform Activation Antibody Sampler Kit: Image 15 Expand Image
    Flow cytometric analysis of Jurkat cells, untreated (blue) or treated with Anisomycin (25µM, 30 min; green) using Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.