Cell Signaling Technology

Product Pathways - MAPK Signaling

Phospho-MAPKAPK-2 (Thr334) (27B7) Rabbit mAb #3007

Applications Reactivity MW (kDa) Source Isotype
W IHC-P IF-IC F H M R Mk 49 Rabbit IgG

Applications Key:  W=Western Blotting  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

Phospho-MAPKAPK-2 (Thr334) (27B7) Rabbit mAb detects endogenous levels of MAPKAPK-2 protein only when phosphorylated at threonine 334.

Source / Purification

Monoclonal antibody is produced by immunizing rabbits with a synthetic phospho-peptide (KLH coupled) corresponding to residues surrounding Thr334 of human MAPKAPK-2.

Western Blotting

Western Blotting

Western blot analysis of extracts from untreated or UV+TPA-treated HeLa and COS cells, using Phospho-MAPKAPK-2 (Thr334) (27B7) Rabbit mAb (upper), or MAPKAPK-2 Antibody #3042 (lower).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, showing nuclear localization, using Phospho-MAPKAPK-2 (Thr334) (27B7) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma untreated (left) or lambda phosphatase-treated (right), using Phospho-MAPKAPK-2 (Thr334) (27B7) Rabbit mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Phospho-MAPKAPK-2 (Thr334) (27B7) Rabbit mAb in the presence of control peptide (left) or Phospho-MAPKAPK-2 (Thr334) Blocking Peptide #1041 (right).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HeLa cells, untreated (blue) or UV-treated (green), using Phospho-MAPKAPK-2 (Thr334) (27B7) Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left), UV-treated (center) or UV and lambda phosphatase-treated (right), using Phospho-MAPKAPK-2 (Thr334) (27B7) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).


Background

In response to cytokines, stress and chemotactic factors, MAP kinase-activated protein kinase 2 (MAPKAPK-2) is rapidly phosphorylated and activated. It has been shown that MAPKAPK-2 is a direct target of p38 MAPK (1). Multiple residues of MAPKAPK-2 are phosphorylated in vivo in response to stress. However, only four residues (Thr25, Thr222, Ser272 and Thr334) are phosphorylated by p38 MAPK in an in vitro kinase assay (2). Phosphorylation at Thr222, Ser272 and Thr334 appears to be essential for the activity of MAPKAPK-2 (2). Thr25 is phosphorylated by p42 MAPK in vitro, but is not required for the activation of MAPKAPK-2 (2).

  1. Rouse, J. et al. (1994) Cell 78, 1027-1037.
  2. Ben-Levy, R. et al. (1995) EMBO J. 14, 5920-5930.

Application References

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Companion Products

Rabbit Monoclonals Produced Using Epitomics® Technology, U.S. Patent No. 5,675,063.

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