Product Pathways - PathScan ELISA
PathScan® Total MEK1 Chemiluminescent Sandwich ELISA Kit #7030
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Important Ordering Details: Product is assembled upon order to ensure maximum activity. Domestic: Please allow up to two weeks for your order to be processed and shipped. International: Please allow up to three weeks, depending on the country, for your order to be processed and shipped.
|Kit Includes||Volume||Solution Color|
|MEK1 mAb Coated Microwells||96 tests|
|MEK1/2 Detection Ab||5.5 ml|
|Anti-rabbit IgG, HRP-linked Antibody||5.5 ml|
|Luminol/Enhancer Solution||3 ml|
|Stable Peroxide Buffer||3 ml|
|Sealing Tape||2 sheets|
|ELISA Wash Buffer (20X)||25 ml|
|Sample Diluent||25 ml|
|Cell Lysis Buffer (10X) #9803||15 ml|
Note: 12 8-well modules –Each module is designed to break apart for 8 tests.
Note: Kit should be stored at 4°C with the exception of Cell Lysis Buffer, which is stored at –20°C (packaged separately).
Species enclosed in parentheses are predicted to react based on 100% sequence homology.
The PathScan® Total MEK1 Chemiluminescent Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total MEK1 protein with a chemiluminescent readout. Chemiluminescent ELISAs often have a wider dynamic range and higher sensitivity than conventional chromogenic detection. This chemiluminescent ELISA, which is offered in low volume microplates, shows increased signal and sensitivity while using smaller sample size. A MEK1 mouse mAb has been coated on the microwells. After incubation with cell lysates, the MEK1 protein is captured by the coated antibody. Following extensive washing, a MEK1/2 rabbit antibody is added to detect the captured total MEK1 protein. Anti-rabbit IgG, HRP-linked antibody is then used to recognize the bound detection antibody. Chemiluminescent reagent is added for signal development. The magnitude of light emission, measured in relative light units (RLU), is proportional to the quantity of total MEK1 protein.
Specificity / Sensitivity
CST's PathScan® Total MEK1 Chemiluminescent Sandwich ELISA Kit #7030 detects endogenous levels of total MEK1 protein in human and mouse cells. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
Relationship between protein concentration of lysates from untreated and PDGF-treated NIH/3T3 cells and immediate light generation with chemiluminescent substrate is shown. Cells (80% confluence) were treated with PDGF (50 ng/ml) and lysed after incubation at 37ºC for 5 minutes. The graph inset corresponding to the shaded area shows high sensitivity and a linear response at the low protein concentration range.
MEK1 and MEK2, also called MAPK or Erk kinases, are dual-specificity protein kinases that function in a mitogen activated protein kinase cascade controlling cell growth and differentiation (1-3). Activation of MEK1 and MEK2 occurs through phosphorylation of two serine residues at positions 217 and 221, located in the activation loop of subdomain VIII, by Raf-like molecules. MEK1/2 is activated by a wide variety of growth factors and cytokines and also by membrane depolarization and calcium influx (1-4). Constitutively active forms of MEK1/2 are sufficient for the transformation of NIH/3T3 cells or the differentiation of PC-12 cells (4). MEK activates p44 and p42 MAP kinase by phosphorylating both threonine and tyrosine residues at sites located within the activation loop of kinase subdomain VIII.
- Crews, C.M. et al. (1992) Science 258, 478-480.
- Alessi, D.R. et al. (1994) EMBO J. 13, 1610-1619.
- Rosen, L.B. et al. (1994) Neuron 12, 1207-1221.
- Cowley, S. et al. (1994) Cell 77, 841-852.
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For Research Use Only. Not For Use In Diagnostic Procedures.