PathScan? Cleaved Notch1 (Val1744) Sandwich ELISA Kit

• 产品编号：CST-7194C      品牌：CST       原厂货号：7194C
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测试剂盒
• 应用分类：

其它组分：

描述：

如图1所示，PathScan® Total Notch1 Sandwich ELISA Kit#7194可以检测1744位缬氨酸位被剪切后的内源Notch1蛋白表达水平。试剂盒的敏感性如图2所示。内部检测结果显示，本试剂盒可以检测如图所示物种的蛋白，同时也可能识别其他物种的同源蛋白。CST公司的PathScan® Cleaved Notch1 (Val1744) Sandwich ELISA Kit是基于“三明治”原理的固相酶联免疫吸附试验（ELISA），用于检测在Val1744位点剪切开的内源性Notch1蛋白。微孔经大鼠来源的Notch1抗体提前包被。加入细胞裂解物孵育后，Notch1蛋白可以被包被的抗体捕获。经过多次冲洗后，加入剪切型Val1744）的兔源Notch1抗体用于检测被捕获到的Notch1蛋白。辣根过氧化物酶（HRP）标记的抗兔IgG抗体被用于检测上述结合的一抗。HRP底物（TMB）用于显色。由此产生的吸光度值与剪切型(Val1744)的Notch1蛋白量成正比。Notch1是一个在细胞发育和命运决定过程中发挥功能的跨膜蛋白（1）。在notch1成熟过程中，notch1分子细胞外结构域会被一个成对碱性氨基酸蛋白酶样的转化酶剪切（2）。在与配体如Delta1结合或是细胞外钙离子耗竭后，notch1的羧基端被释放并随后在Gly1743 和 Val1744之间被切开（3,4）。结果导致细胞内处于活化状态的notch1片段转运至细胞核内并转录激活。

1. Artavanis-Tsakonas, S. et al. (1999) Science 284, 770-6.
2. Chan, Y.M. and Jan, Y.N. (1998) Cell 94, 423-6.
3. Schroeter, E.H. et al. (1998) Nature 393, 382-6.
4. Rand, M.D. et al. (2000) Mol Cell Biol 20, 1825-35.

原厂资料：

Specificity / Sensitivity

PathScan® Cleaved Notch1 (Val1744) Sandwich ELISA Kit #7194 detects endogenous levels of Notch1 protein when cleaved at Val1744 as shown in Figure 1. The 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

PathScan® Cleaved Notch1 (Val1744) Sandwich ELISA Kit from Cell Signaling Technology is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Notch1 when cleaved at Val1744. A Notch1 rat antibody has been coated onto the microwells. After incubation with cell lysates, Notch1 protein is captured by the coated antibody. Following extensive washing, a cleaved Notch1 (Val1744) Rabbit detection antibody is added to detect the captured Notch1 protein. Anti-Rabbit IgG, HRP-linked Antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of cleaved Notch1 (Val1744).

Background

Notch1 is a transmembrane protein functioning in development and the determination of cell fate (1). During maturation, the notch molecule is cleaved by a furin-like convertase at its extracellular domain (2). Upon binding to a ligand such as Delta1, or upon extracellular calcium depletion, the carboxy-terminal notch1 fragment is released and further cleaved between Gly1743 and Val1744 (3,4). The resulting activated cytosolic fragment translocates to the nucleus where it activates transcription.

1. Artavanis-Tsakonas, S. et al. (1999) Science 284, 770-6.
2. Chan, Y.M. and Jan, Y.N. (1998) Cell 94, 423-6.
3. Schroeter, E.H. et al. (1998) Nature 393, 382-6.
4. Rand, M.D. et al. (2000) Mol Cell Biol 20, 1825-35.

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).