p38 MAPK

GWB-6BD47E

1432

Synthetic phosphopeptide corresponding to an amino acid sequence within p38 MAPK which includes phosphorylated Thr180 and Tyr182.

Antisera to phosphorylated rat p38 MAPK were raised by repeated immunisations of rabbits with highly purified antigen. Purified IgG prepared by affinity chromatography.

Western Blotting - 1:1000Protocols from PublicationsCitation: 1: Chen D Wilkinson CR Watt S Penkett CJ Toone WM Jones N Bä hler J. Multiple pathways differentially regulate global oxidative stress responses in fission yeast. Mol Biol Cell. 2008 Jan; 19(1):308-17. Epub 2007 Nov 14. PubMed PMID: 18003976; PubMed Central PMCID: PMC2174203. Experiment Name: Western analyses of protein extracts from TBH and HP stress-time course experimentsExperiment Background: Genome-wide analyses have uncovered hundreds of genes whose expression is modulated by Sty1p and Atf1p in response to HP (hydrogen peroxide) and other stressors like TBH (t-butylhydroperoxide). Sty1p became strongly phosphorylated in HP (hydrogen peroxide) but less so in TBH (t-butylhydroperoxide). The fission yeast Schizosaccharomyes pombe is a popular model organism to study oxidative stress response pathways. Experiment Steps: 1. For stress experiments protein extract preparation grow cells in yeast extract (YE) medium at 30C to a titer of ~2 X 106 cells/ml. 2. Add hydrogen peroxide (HP) to a final concentration of 0. 07 0. 5 or 6 mM; and add tert-butylhydroperoxide to a final concentration of 2 mM. 3. Harvest cells by filtration before and after treatment and freeze on liquid nitrogen. 4. Thaw the filters in STOP buffer (150 mM NaCl 50 mM NaF 10 mM EDTA and 1 mM NaN3 pH 8) and wash off cells and harvest cell by centrifugation (2000 rpm for 2 min). 5. Resuspend cells in lysis buffer [The composition of the lysis buffer is 50 mM HEPES pH 7. 5 40 mM b-glycerophosphate 1 mM sodium vanadate 50 mM NaF 0. 5% NP-40 150 mM NaCl 1 mM phenylmethylsulfonyl fluoride and 1 complete mini protease inhibitor tablet. ]6. After glass bead lysis boil the supernatant in 2X SDS loading buffer. 7. Separate extracts (50 micro gram) by SDS-polyacrylamide gel electrophoresis on a 10% gel and8. Analyze by western blotting with anti-phospho p38 ( San Diego CA) or anti-hemagglutinin (HA) diluted at 1:1000.

ATP + a protein = ADP + a phosphoprotein. Cofactor: Magnesium.

Binds to a kinase interaction motif within the protein tyrosine phosphatase PTPRR. This interaction retains MAPK14 in the cytoplasm and prevents nuclear accumulation. Interacts with SPAG9 (By similarity). Interacts with NP60 and FAM48A.

Brain heart placenta pancreas and skeletal muscle. Expressed to a lesser extent in lung liver and kidney.

Dually phosphorylated on Thr-180 and Tyr-182 which activates the enzyme.

Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. MAP kinase subfamily.