Cell Signaling Technology

Product Pathways - PathScan ELISA

PathScan® Phospho-MEK1 (Ser217/221) Chemiluminescent Sandwich ELISA Kit #7029

When ordering five or more kits, please contact us for processing time and pricing at sales@cellsignal.com.

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 (61B12) mAb Coated Microwells 96 tests
P-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
ELISA Sample Diluent 25 ml Blue
Cell Lysis Buffer (10X) #9803 15 ml Yellowish

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 Cross-Reactivity

H M

Reactivity Key:  H=Human  M=Mouse
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Description

The PathScan® Phospho-MEK1 (Ser217/221) Chemiluminescent Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of phospho-MEK1 (Ser217/221) protein with a chemiluminescent readout. Chemiluminescence 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 onto the microwells. After incubation with cell lysates, total MEK1 protein (phospho- and nonphospho-) is captured by the coated antibody. Following extensive washing, a phospho-MEK1/2 (Ser217/221) rabbit antibody is added to detect the captured phospho-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 phospho-MEK1 (Ser217/221) protein.Antibodies in kit are custom formulations specific to kit.

Specificity / Sensitivity

PathScan® Phospho-MEK1 (Ser217/221) Chemiluminescent Sandwich ELISA Kit #7029 detects endogenous levels of phospho-MEK1 (Ser217/221) 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.

ELISA

ELISA

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. Graph inset corresponding to the shaded area shows high sensitivity and a linear response at the low protein concentration range.

Background

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.

  1. Crews, C.M. et al. (1992) Science 258, 478-480.
  2. Alessi, D.R. et al. (1994) EMBO J. 13, 1610-1619.
  3. Rosen, L.B. et al. (1994) Neuron 12, 1207-1221.
  4. Cowley, S. et al. (1994) Cell 77, 841-852.

Application References

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Protocols

* Product-specific protocol.

Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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