Product Pathways - PathScan ELISA
PathScan® Phospho-p44/42 MAPK (Thr202/Tyr204) Sandwich ELISA Antibody Pair #7246
|7246S||1 Kit (Reagents for 4 x 96 well plates)||---||In Stock||---|
|7246||carrier free and custom formulation / quantity||email request|
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|Kit Includes||Volume||Cap Color|
|P-p44/42 MAPK (T202/ Y204) Capture Ab (100X)||0.4 ml||Pink|
|p44/42 MAPK DetectionAb (100X)||0.4 ml||Blue|
|Anti-mouse IgG, HRP-linked Antibody (1000X)||0.04 ml||Yellow|
Storage: Capture and detection antibodies are stored at 4°C. HRP-linked secondary reagent is stored at -20°C.
CST's PathScan® Phospho-p44/42 MAPK (Thr202/Tyr204) Sandwich ELISA Antibody Pair is offered as an economical alternative to our PathScan® Phospho-p44/42 MAPK (Thr202/Tyr204) Sandwich ELISA Kit #7177. Capture and detection antibodies (100X stocks) and an HRP-conjugated secondary antibody (1000X stock) are supplied. Sufficient reagents are supplied for 4 x 96 well ELISAs. The Phospho-p44/42 MAPK Rabbit Capture Antibody is coated in PBS overnight in a 96 well microplate. After blocking, cell lysate is added followed by a p44/42 MAPK Mouse Detection Antibody and an HRP-conjugated Anti-Mouse IgG Antibody. HRP substrate, TMB, is added for color development. The magnitude of the absorbance for this developed color is proportional to the quantity of p44/42 MAPK phosphorylated at Thr202/Tyr204.
Antibodies in kit are custom formulations specific to kit.
Specificity / Sensitivity
For Antibody Pair specificity and sensitivity, please refer to the corresponding PathScan® Sandwich ELISA Kit. Note: This antibody pair detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
The relationship between the protein concentration of the lysate from untreated and PDGF-treated NIH/3T3 cells and the absorbance at 450 nm using PathScan® Phospho-p44/42 MAPK (Thr202/Tyr204) Sandwich ELISA Antibody Pair #7246 is shown. NIH/3T3 cells were treated with PDGF (100 ng/mL) for 5 minutes at 37ºC and then lysed.
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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- 9803 Cell Lysis Buffer (10X)
- 7004 TMB Substrate
- 7002 STOP Solution
- 9808 Phosphate Buffered Saline (PBS-20X)
- 9809 Phosphate Buffered Saline with Tween® 20 (PBST-20X)
For Research Use Only. Not For Use In Diagnostic Procedures.
PathScan® is a trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.