Western blot analysis of extracts from HeLa cells, untreated (-) or UV- and TPA-treated (+), using Phospho-MAPKAPK-2 (Thr222) (9A7) Rabbit mAb (upper), or MAPKAPK-2 Antibody #3042 (lower).
Immunoprecipitation of MAPKAPK-2 from HeLa cell extracts using MAPKAPK-2 (D1E11) Rabbit mAb (lane 2), MAPKAPK-2 Antibody #3042 (lane 3), or Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 4). Lane 1 is 10% input. Western blot analysis was performed using MAPKAPK-2 (D1E11) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb #3678 was used as a secondary antibody.
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
Western blot analysis of extracts from NIH/3T3 cells, untreated (-) or UV- and TPA-treated (+), using Phospho-MAPKAPK-2 (Thr222) (9A7) Rabbit mAb (upper) or MAPKAPK-2 Antibody #3042 (lower).
Western blot analysis of extracts from various cell lines, untreated (-) or treated with UV (40 mJ/cm2, 30 min recovery; +), using MAPKAPK-2 (D1E11) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
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).
Western blot analysis of cell extracts from MAPKAPK-2 wild type (WT) and knockout (KO) mouse embryonic fibroblasts (MEF) using MAPKAPK-2 (D1E11) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). MAPKAPK-2 WT and KO MEFs were generously provided by Drs. Sandra Morandell and Michael Yaffe, David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA.
|Phospho-MAPKAPK-2 (Thr334) (27B7) Rabbit mAb 3007||20 µl||
||H Mk M R||49||Rabbit IgG|
|Phospho-MAPKAPK-2 (Thr222) (9A7) Rabbit mAb 3316||20 µl||
||H Mk M R||49||Rabbit IgG|
|MAPKAPK-2 (D1E11) Rabbit mAb 12155||20 µl||
||B H Mk M Pg R||47, 49 (Phosphorylated)||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody 7074||100 µl||
The MAPKAPK-2 Antibody Sampler Kit provides an economical means of evaluating levels of MAPKAPK-2 protein phosphorylated at specific sites, as well as total MAPKAPK-2 protein levels. The kit contains enough primary and secondary antibodies to perform two Western blot experiments.
Each antibody in the MAPKAPK-2 Antibody Sampler Kit detects endogenous levels of its target protein. Activation state antibodies detect only target proteins phosphorylated at indicated residues. MAPKAPK-2 (D1E11) Rabbit mAb is not known to cross-react with other MAPKAPK proteins.
Monoclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Thr222 and Thr334 of human MAPKAPK-2 and nonphospho peptides corresponding to residues surrounding Pro381 of human MAPKAPK-2 protein..
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).
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