Cell Signaling Technology

Product Pathways - PathScan ELISA

PathScan® Total MEK1 Sandwich ELISA Antibody Pair #7215

Kit Includes Volume Cap Color
Total MEK1 Capture Antibody (100X) 0.4 milliliters Pink
Total MEK1/2 Detection Antibody (100X) 0.4 milliliters Blue
Anti-Rabbit IgG, HRP-LInked Antibody (1000X) 0.04 milliliters Red

Capture and Detection Antibodies are stored at 4°C. HRP-Linked Secondary Antibody is stored at -20°C.

Species Cross-Reactivity

H M

Reactivity Key:  H=Human  M=Mouse

Description

CST's PathScan® Total MEK1 Sandwich ELISA Antibody Pair is being offered as an economical alternative to our PathScan® Total MEK1 Sandwich ELISA Kit #7165. Capture and Detection antibodies (100X stocks) and HRP-conjugated secondary antibody (1000X stock) are supplied. Sufficient reagents are supplied for 4 x 96 well ELISAs. The MEK1 Capture Antibody is coated in PBS overnight in a 96 well microplate. After blocking, cell lysates are added followed by a MEK1/2 Detection Antibody and anti-Rabbit IgG, HRP-conjugated antibody. HRP substrate, TMB, is added for color development. The magnitude of the absorbance for this developed color is proportional to the quantity of total MEK1/2 protein.*Antibodies in this kit are custom formulations specific to the kit.

Sandwich ELISA

Sandwich ELISA

The relationship between lysate protein concentration from untreated and PDGF-treated NIH/3T3 cells and the absorbance at 450 nm using PathScan® Total MEK1 Sandwich ELISA Antibody Pair #7215 is shown. NIH/3T3 cells were treated with PDGF for 20 minutes at 37°C and then lysed.

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 (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 PC12 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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