Cell Signaling Technology
XP Monoclonal Antibody

Product Pathways - MAPK Signaling

MSK2 (D41A4) XP® Rabbit mAb #3679

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IF-IC H Endogenous 85, 90 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

MSK2 (D41A4) XP® Rabbit mAb detects endogenous levels of total MSK2 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the region surrounding Pro751 of human MSK2.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa and A-431 cells, untreated or treated with Calyculin A #9902 (100 nM, 30 minutes), using MSK2 (D41A4) XP® Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of A-431 cells using MSK2 (D41A4) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Background

Mitogen- and stress-activated protein kinase 1 (MSK1) and MSK2 are serine/threonine kinases that promote immediate early gene transcription in stress- or mitogen-induced cells (1-4,7, 8) and LPS-stimulated macrophages (9). MSK2, also known as RSKB, contains two catalytic domains and has been shown to interact directly with p38 MAP kinase (10). MSK2 is phosphorylated and activated in response to tumor necrosis factor, epidermal growth factor or phorbol ester in HeLa cells or murine embryonic fibroblasts (MEFs) in a p38- and ERK-dependent manner (8,11). Phosphorylation on residues Ser196 and Thr568 within the activation loop of both catalytic domains is required for full kinase activation (11). Both MSK1 and MSK2 contain a functional nuclear localization sequence that is sufficient and required for nuclear targeting (10). Consistent with their nuclear localization, these kinases play an important role in regulating transcriptional responses to stress and mitogens. Activation of MSK2 in HeLa cells or MEFs results in rapid phosphorylation of histone H3, HMG-14, CREB and ATF1 and acetylation of histone H3 associated with immediate early gene transcription (3,4,6,7).

  1. Ananieva, O. et al. (2008) Nat Immunol 9, 1028-36.
  2. Sury, M.D. et al. (2006) Free Radic Biol Med 41, 1372-83.
  3. Duncan, E.A. et al. (2006) J Biol Chem 281, 12521-5.
  4. Darragh, J. et al. (2005) Biochem J 390, 749-59.
  5. Doehn, U. et al. (2004) Biochem J 382, 425-31.
  6. Davie, J.R. (2003) Sci STKE 2003, PE33.
  7. Soloaga, A. et al. (2003) EMBO J 22, 2788-97.
  8. Wiggin, G.R. et al. (2002) Mol Cell Biol 22, 2871-81.
  9. Caivano, M. and Cohen, P. (2000) J Immunol 164, 3018-25.
  10. Tomás-Zuber, M. et al. (2001) J Biol Chem 276, 5892-9.
  11. Tomás-Zuber, M. et al. (2000) J Biol Chem 275, 23549-58.

Application References

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Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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