MEK1/2 Control Cell Extracts

• 产品编号：CST-9160S      品牌：CST       原厂货号：9160S
• 产品分类：蛋白质表达与纯化 > 蛋白质样本制备 > 细胞组织粗提物与裂解物
• 应用分类：

描述：

非磷酸化的MEK1/2 细胞提取物: 未经处理的HeLa细胞总提取物，可以作为阴性对照。细胞提取物以SDS样品缓冲液的形式提供。保存于-20ºC。磷酸化的MEK1/2细胞提取物:用TPA (200 nM, 15 min)处理的HeLa细胞总提取物，可以作为阳性对照。细胞提取物以SDS样品缓冲液的形式提供。保存于-20ºC。Western Blots：作为对照，我们建议使用20 µl磷酸化和非磷酸化的MEK1/2对照提取物。在使用之前煮沸。MEK1、MEK2,也被称为MAPK或Erk激酶，它们是双特异性蛋白激酶，在丝裂原活化蛋白激酶级联反应中发挥作用，控制细胞生长和分化(1-3)。MEK1、MEK2的活化是通过Raf样分子对两个丝氨酸（217 /221位点）的磷酸化而实现的，这两个丝氨酸位点位于亚结构域VIII的活化环内。多种生长因子和细胞因子以及膜去极化和钙内流都能激活MEK1/2(1-4)。组成型活化的MEK1/2能够诱导NIH/3T3细胞的转化和PC-12细胞的分化(4)。MEK蛋白通过将p44、p42 MAPK激酶上位于亚结构域VIII的活化环内的丝氨酸和苏氨酸磷酸化，激活p44、 p42 MAPK激酶。

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.

原厂资料：

Description

Nonphosporylated MEK1/2 Cell Extracts: Total cell extracts from HeLa cells prepared without treatment, serve as a negative control. Supplied in SDS Sample Buffer. Store at -20ºC. 
 
 Phosphorylated MEK1/2 Cell Extracts: Total cell extracts from HeLa cells prepared with TPA treatment (200 nM, 15 min) serve as a positive control. Supplied in SDS Sample Buffer. Store at -20ºC.

Applications

Western Blots: As controls, we recommend using 20 µl of phosphorylated and nonphosphorylated MEK1/2 control extracts. Boil sample before use.

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.

注意事项：

Storage: Supplied in SDS Sample Buffer: 62.5 mM Tris-
HCL (pH 6.8 at 25°C), 2% w/v SDS, 10% glycerol, 50 mM
DTT, 0.01% w/v bromophenol blue or phenol red. Store at
–20°C.