MEK1 Total, l Sandwich ELISA Kit, BioAssay (Map Kinase Kinase, MAPK Kinase, Erk Kinase 1)

Detects endogenous levels of total MEK1 protein. A significant induction of MEK1 phosphorylation can be detected in PMA-treated NIH/3T3 cells using Phospho- MEK1 (Ser217/221) Sandwich ELISA Kit. However, the level of total MEK1 detected by this sandwich ELISA kit remains unchanged. This kit can also be used to detect total MEK1 protein in human 293 cells. Total MEK1 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total MEK1 protein. 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, MEK1/2 Antibody is added to detect the captured MEK1 protein. HRP-linked anti-rabbit antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of optical density for this developed color is proportional to the quantity of total, MEK1 protein. 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. Specificity: Human and mouse, Kit Contents: MEK1 Mouse mAb coated Microwells 96 tests, MEK1/2 Detection Ab 11ml green, Anti-rabbit IgG HRP-Linked Ab 11ml red, TMB Substrate 11ml colorless, STOP Solution 11ml colorless, 20X Wash Buffer 25ml colorless, Sample Diluent 25ml blue, 10X Cell Lysis Buffer 15ml yellowish, Storage and Stability: Store kit components at 4°C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.