PathScan? Phospho-MARCKS (Ser152/156) Sandwich ELISA Kit

• 产品编号：CST-7251C      品牌：CST       原厂货号：7251C
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测试剂盒
• 应用分类：

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描述：

如图1所示，夹心酶联免疫吸附测定试剂盒PathScan® Phospho-MARCKS (Ser152/156) Sandwich ELISA Kit＃7251检测内源性丝氨酸152和156位点磷酸化的MARCKS蛋白。试剂盒灵敏度如图2所示。本试剂盒用来检测指定物种的蛋白，即通过内部测试确定的指定物种蛋白，也可能可以用来检测来自其他物种的同源蛋白。PathScan® 磷酸-MARCKS（Ser152/156）夹心酶联免疫吸附测定试剂盒是一个固相夹心酶联免疫吸附试验（ELISA），检测内源性的丝氨酸152和156位磷酸化的MARCKS蛋白。MARCKS兔抗体包被微孔板。细胞裂解物孵育后，MARCKS（磷酸和非磷酸）由包被的抗体捕获。充分洗涤后，添加生物素化的磷酸-MARCKS（Ser152/156）兔单克隆以检测捕获的磷酸化-MARCKS蛋白。然后使用HRP-标记的链霉抗生物素蛋白检测识别结合的抗体。添加HRP底物TMB，显色。显色的吸光度大小与MARCKS磷酸化丝氨酸152和156位的含量成正比。试剂盒中的抗体是特别针对试剂盒定制的。豆寇酰化富含丙氨酸C-激酶底物（MARCKS）是一种重要的PKC底物，在多种细胞类型中表达。 MARCKS与细胞活力，细胞粘附，吞噬，膜运输和有丝分裂有关（1）。PKC响应生长因子和氧化应激磷酸化MARCKS的丝氨酸（159，163，167和170位），调节钙/钙调蛋白结合和MARCKS丝状（F）-肌动蛋白交联反应活性（2-4）。PKC磷酸化作用导致MARCKS从细胞膜到细胞质迁移(5)。

1. Ramsden, J.J. (2000) Int J Biochem Cell Biol 32, 475-9.
2. Heemskerk, F.M. et al. (1993) Biochem Biophys Res Commun 190, 236-41.
3. Graff, J.M. et al. (1989) J Biol Chem 264, 21818-23.
4. Hartwig, J.H. et al. (1992) Nature 356, 618-22.
5. Thelen, M. et al. (1991) Nature 351, 320-2.

原厂资料：

Specificity / Sensitivity

PathScan® Phospho-MARCKS (Ser152/156) Sandwich ELISA Kit #7251 detects endogenous levels of MARCKS protein only when phosphorylated at Serines 152 and 156 as shown in Figure 1. Kit sensitivity is shown in Figure 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Description

The PathScan® Phospho-MARCKS (Ser152/156) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of MARCKS when phosphorylated at Serines 152 and 156. A MARCKS rabbit antibody has been coated onto the microwells. After incubation with cell lysates, MARCKS protein (phosphorylated and nonphospho) is captured by the coated antibody. Following extensive washing, a biotinylated phospho-MARCKS (Ser152/156) rabbit monoclonal detection antibody is added to detect the captured phospho-MARCKS protein. HRP-linked streptavidin is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for the developed color is proportional to the quantity of MARCKS phosphorylated at Serines 152 and 156. 
 Antibodies in kit are custom formulations specific to kit.

Background

Myristoylated Alanine-Rich C-Kinase Substrate (MARCKS) is a major PKC substrate expressed in many cell types. MARCKS has been implicated in cell motility, cell adhesion, phagocytosis, membrane traffic, and mitogenesis (1). PKC phosphorylates Ser159, 163, 167, and 170 of MARCKS in response to growth factors and oxidative stress. Phosphorylation at these sites regulates the calcium/calmodulin binding and filamentous (F)-actin cross-linking activities of MARCKS (2-4). Phosphorylation by PKC also results in translocation of MARCKS from the plasma membrane to the cytoplasm (5).

1. Ramsden, J.J. (2000) Int J Biochem Cell Biol 32, 475-9.
2. Heemskerk, F.M. et al. (1993) Biochem Biophys Res Commun 190, 236-41.
3. Graff, J.M. et al. (1989) J Biol Chem 264, 21818-23.
4. Hartwig, J.H. et al. (1992) Nature 356, 618-22.
5. Thelen, M. et al. (1991) Nature 351, 320-2.

注意事项：

* 12 8-well modules -Each module is designed to break apart for 8 tests.
**Kit should be stored at 4°C with the exception of 10X Cell Lysis Buffer, which is stored at –20°C (packaged separately).