Product Pathways - MAPK Signaling
Phospho-Erk1/2 Pathway Sampler Kit #9911
|9911S||1 Kit (5 x 40 µl)||---||In Stock||---|
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|Kit Includes||Quantity||Applications||Reactivity||Homology†||MW (kDa)||Isotype|
|Phospho-c-Raf (Ser338) (56A6) Rabbit mAb #9427||40 µl||W||H, M, R, Mk||74||Rabbit IgG|
|Phospho-MEK1/2 (Ser217/221) (41G9) Rabbit mAb #9154||40 µl||W, IP||H, M, R, Mk, Dm||C||45||Rabbit|
|Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370||40 µl||W, IP, IHC-P, IF-IC, F||H, M, R, Hm, Mk, Mi, Dm, Z, B, Dg, Pg, Sc||C, Ce||44, 42||Rabbit IgG|
|Phospho-p90RSK (Ser380) (9D9) Rabbit mAb #9335||40 µl||W||H, M, R, Mk||C||90||Rabbit IgG|
|Phospho-MSK1 (Thr581) Antibody #9595||40 µl||W, IP, IHC-P||H, M||90||Rabbit|
|Anti-rabbit IgG, HRP-linked Antibody #7074||100 µl||Goat|
|U0126 #9903||.3 mg||380.50|
†Species predicted to react based on 100% sequence homology.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry
Reactivity Key: H=Human, M=Mouse, R=Rat, Mk=Monkey, Dm=D. melanogaster, Hm=Hamster, Mi=Mink, Z=Zebrafish, B=Bovine, Dg=Dog, Pg=Pig, Sc=S. cerevisiae
Western blot analysis of extracts from 293, NIH/3T3 and C6 cells, treated with λ phosphatase or TPA as indicated, using Phospho-p44/42 MAPK (Thr202/Tyr204) (D13.14.4E) Rabbit mAb #4370 (upper), or p44/42 MAP Kinase (137F5) Rabbit mAb #4695 (lower).
Western blot analysis of extracts from untreated or TPA-treated 293 and NIH/3T3 cells using Phospho-p90RSK (Ser380) (9D9) Rabbit mAb #9335.
Western blot analysis of extracts from NIH/3T3, HeLa and COS cells, untreated or treated with TPA, using Phospho-c-Raf (Ser338) (56A6) Rabbit mAb #9427.
Western blot analysis of extracts from 293 cells untreated or treated with UV and HeLa cells untreated or treated with calyculin A using Phospho-MSK1 (Thr581) Antibody #9595.
Western blot analysis of extracts from untreated or TPA-treated HeLa and NIH/3T3 cells, using Phospho-MEK1/2 (Ser217/221) (41G9) Rabbit mAb #9154 (upper), or MEK1/2 Antibody #9122 (lower).
Immunohistochemical analysis of paraffin-embedded human lung carcinoma, untreated (left) or λ phosphatase-treated (right), using Phospho-p44/42 MAPK (Thr202/Tyr204) (D13.14.4E) Rabbit mAb #4370.
Flow cytometric analysis of Jurkat cells, U0126-treated (blue) or PMA-treated (green), using Phospho-p44/42 MAPK (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370.
Confocal immunofluorescent analysis of C2C12 cells, U0126-treated (left, #9903 10 μM for 1 h) or TPA-treated (right, #9905 200 nM for 15 min), using Phospho-p44/42 MAPK (Thr202/Tyr204) (D13.14.4E) Rabbit mAb #4370 (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).
The Phospho-Erk1/2 Pathway Sampler Kit provides an economical means of evaluating multiple members of the Erk pathway as well as their activation state. The kit contains enough primary and secondary antibodies to perform four Western blot experiments.
Specificity / Sensitivity
Each antibody in the Phospho-Erk1/2 Pathway Sampler Kit recognizes only the phosphorylated form of its specific target.
Source / Purification
Polyclonal and monoclonal antibodies are produced by immunizing rabbits with synthetic phosphopeptides corresponding to residues surrounding Ser338 of human Raf, Ser217/221 of human MEK1/2, Thr202/Tyr204 of human p44 MAP kinase, Ser380 of human p90RSK or Thr581 of human MSK1. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.
Mitogen-activated protein kinases (MAPKs) are a widely conserved family of serine/threonine protein kinases involved in many cellular programs, such as cell proliferation, differentiation, motility, and death. The p44/42 MAPK (Erk1/2) signaling pathway can be activated in response to a diverse range of extracellular stimuli including mitogens, growth factors, and cytokines (1-3), and research investigators consider it an important target in the diagnosis and treatment of cancer (4). Upon stimulation, a sequential three-part protein kinase cascade is initiated, consisting of a MAP kinase kinase kinase (MAPKKK or MAP3K), a MAP kinase kinase (MAPKK or MAP2K), and a MAP kinase (MAPK). Multiple p44/42 MAP3Ks have been identified, including members of the Raf family, as well as Mos and Tpl2/COT. MEK1 and MEK2 are the primary MAPKKs in this pathway (5,6). MEK1 and MEK2 activate p44 and p42 through phosphorylation of activation loop residues Thr202/Tyr204 and Thr185/Tyr187, respectively. Several downstream targets of p44/42 have been identified, including p90RSK (7) and the transcription factor Elk-1 (8,9). p44/42 are negatively regulated by a family of dual-specificity (Thr/Tyr) MAPK phosphatases, known as DUSPs or MKPs (10), along with MEK inhibitors, such as U0126 and PD98059.
- Roux, P.P. and Blenis, J. (2004) Microbiol Mol Biol Rev 68, 320-44.
- Baccarini, M. (2005) FEBS Lett 579, 3271-7.
- Meloche, S. and Pouysségur, J. (2007) Oncogene 26, 3227-39.
- Roberts, P.J. and Der, C.J. (2007) Oncogene 26, 3291-310.
- Rubinfeld, H. and Seger, R. (2005) Mol Biotechnol 31, 151-74.
- Murphy, L.O. and Blenis, J. (2006) Trends Biochem Sci 31, 268-75.
- Dalby, K.N. et al. (1998) J Biol Chem 273, 1496-505.
- Marais, R. et al. (1993) Cell 73, 381-93.
- Kortenjann, M. et al. (1994) Mol Cell Biol 14, 4815-24.
- Owens, D.M. and Keyse, S.M. (2007) Oncogene 26, 3203-13.
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* Product-specific protocol.
- 9160 MEK1/2 HeLa Control Cell Extracts
- 9126 MEK1/2 (47E6) Rabbit mAb
- 4695 p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb
- 9355 RSK1/RSK2/RSK3 (32D7) Rabbit mAb
- 9422 c-Raf Antibody
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
For Research Use Only. Not For Use In Diagnostic Procedures.
DRAQ5® is a registered trademark of Biostatus Limited.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
Select rabbit monoclonal antibodies are developed, validated, and produced at CST using in part technology under license (granting certain rights including those under U.S. Patents No. 5,675,063 and in some instances 7,429,487) from Epitomics, Inc.