Cell Signaling Technology

Product Pathways - MAPK Signaling

p38 MAPK Isoform Activation Antibody Sampler Kit #9790

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
p38α MAPK Antibody #9218 40 µl W IP IHC-P H M R Mk 40 Rabbit
p38β MAPK (C28C2) Rabbit mAb #2339 40 µl W IP H Mk 43 Rabbit IgG
p38γ MAPK Antibody #2307 40 µl W IP H M R Mk 46 Rabbit
p38δ MAPK (10A8) Rabbit mAb #2308 40 µl W IP H R 43 Rabbit IgG
Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb #4511 40 µl W IP IHC-P IF-IC F H M R Mk Mi Pg Sc (Hm) (C) (Z) (B) 43 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  C=Chicken  Mi=Mink  Z=Zebrafish  B=Bovine  Pg=Pig  Sc=S. cerevisiae
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

Each isoform-specific antibody in the p38 MAPK Isoform Activation Antibody Sampler Kit recognizes the appropriate isoform of p38 MAPK. The Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb recognizes all four isoforms of p38 MAPK when phosphorylated at the specified sites.

Western Blotting

Western Blotting

Western blot analysis of purified recombinant full-length p38 MAP kinase GST fusion proteins using p38 MAP kinase pan antibody (upper) or p38γ MAP Kinase Antibody #2307 (lower).

Western Blotting

Western Blotting

Western blot analysis of purified recombinant full-length p38 MAP kinase GST fusion proteins using p38 MAP kinase pan antibody (upper), or p38δ MAP Kinase (10A8) Rabbit mAb #2308 (lower).

Western Blotting

Western Blotting

Western blot analysis of purified recombinant full-length p38 MAP kinase GST-fusion proteins using p38β MAP Kinase (C28C2) Rabbit mAb #2339 (upper), or a p38 MAP kinase pan antibody (lower).


Western Blotting

Western Blotting

Western blot analysis of extracts from COS and 293 cells, untreated or UV-treated, using Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb #4511 (upper) or p38 MAPK Antibody #9212 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from Xenopus oocytes overexpressing Myc-tagged p38 isoforms using p38α MAPK Antibody #9218 (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.)

IP

IP

Immunoprecipitation of phospho-p38 from THP1 cells, untreated or sorbitol-treated, using phospho-p38 MAPK (Thr180/Tyr182) Antibody #4511, followed by western blot with p38γ Antibody #2307 (right two lanes). Left two lanes are THP1 whole cell extracts representing 10% of the IP input. Western blot was detected using the Mouse Anti-rabbit IgG (Confirmation Specific) mAb #3678.


IP

IP

Immunoprecipitation of phospho-p38 from 293 cells, untreated, sorbitol or anisomycin-treated, using phospho-p38 MAPK (Thr180/Tyr182) Antibody #4511, followed by western blot with p38δ Antibody #2308 (right three lanes). Left three lanes are 293 whole cell extracts representing 10% of the IP input. Western blot was detected using the Mouse Anti-rabbit IgG (Confirmation Specific) mAb #3678.

IP

IP

Immunoprecipitation of phospho-p38 from 293 cells, untreated, sorbitol or anisomycin-treated, using phospho-p38 MAPK (Thr180/Tyr182) Antibody #4511, followed by western blot with p38β Antibody #2339 (right three lanes). Left three lanes are 293 whole cell extracts representing 10% of the IP input. Western blot was detected using the Mouse Anti-rabbit IgG (Confirmation Specific) mAb #3678.

IP

IP

Immunoprecipitation of phospho-p38 from THP1 cells, untreated or sorbitol-treated, using phospho-p38 MAPK (Thr180/Tyr182) Antibody #4511, followed by western blot with p38α Antibody #9218 (right two lanes). Left two lanes are THP1 whole cell extracts representing 10% of the IP input.


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 four IP/western blot experiments.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to carboxy-terminal residues of human p38α MAPK or human p38γ MAPK. Antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding the carboxy terminus of p38β MAPK or p38δ MAPK and 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, 1027-1037.
  2. Han, J. et al. (1994) Science 265, 808-811.
  3. Lee, J.C. et al. (1994) Nature 372, 739-746.
  4. Freshney, N.W. et al. (1994) Cell 78, 1039-1049.
  5. Raingeaud, J. et al. (1995) J. Biol. Chem. 270, 7420-7426.
  6. Zervos, A.S. et al. (1995) Proc. Natl. Acad. Sci. USA 92, 10531-10534.
  7. Zhao, M. et al. (1999) Mol. Cell. Biol. 19, 21-30.
  8. Yang, S.H. et al. (1999) Mol. Cell. Biol. 19, 4028-4038.
  9. Cuenda, A. et al. (1995) FEBS Lett 364, 229-33.
  10. Kumar, S. et al. (1999) Biochem Biophys Res Commun 263, 825-31.

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