Science Catalog 2012 Life Cell Signaling A. Anti-ACTIVE® MAPK kDa – + NGF 98 – 64 – 50 – 36 – 1 2 B. Anti-MAPK kDa – + NGF 98 – 64 – 50 – 36 – 1 2 3 4 5 6 Detection of MAPK, JNK and p38 in PC12 cell extracts. Panel A. Western blot analysis using Anti-ACTIVE® MAPK, Anti-ACTIVE® JNK and Anti-ACTIVE® p38 polyclonal antibodies to detect activated MAPK, JNK and p38. Panel B. Western blot analysis using anti-MAPK, anti-JNK and anti-p38 antibodies to detect activated and nonactivated MAPK, JNK and p38 in untreated or NGF- or sorbitol-treated PC12 cells. Detection of activated MAPK, JNK and p38 in PC12 cells by immunocytochemistry. PC12 cells were grown to 80% confluence in RPMI 1640 medium supplemented with 25mM HEPES, 300mg/L l-glutamine, 10% horse serum, 5% fetal bovine serum and 0.5mM EGTA. Cells were either untreated or treated with 200ng/ml NGF or 1M sorbitol as indicated. ICC was performed as described in Promega Technical Bulletin #TB262. Anti-ACTIVE® antibodies were used at the following dilutions: Panel A. MAPK, 1:500; Panel B. JNK, 1:1,000; Panel C. p38, 1:500. Protocols developed and performed at Promega. 3 45 6 Anti-JNK – + Sorbitol Anti-p38 – + Sorbitol Anti-ACTIVE® JNK – + Sorbitol Anti-ACTIVE® p38 – + Sorbitol A. Anti-ACTIVE® MAPK pAb NGF-Treated Untreated – ERK1 – ERK2 – JNK2 – JNK1 – p38 B. Anti-ACTIVE® JNK pAb Sorbitol-Treated Untreated – ERK1 – ERK2 – JNK2 – JNK1 – p38 C. Anti-ACTIVE® p38 pAb Sorbitol-Treated Untreated 168 For complete and up-to-date product information visit: www.promega.com/catalog 1859TC 2870TA02_0A Pagina 171
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