PathScan? Phospho-c-Abl (panTyr) Sandwich ELISA Kit

• 产品编号：CST-7903C      品牌：CST       原厂货号：7903C
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测试剂盒
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描述：

当人细胞中酪氨酸残基磷酸化时，PathScan® Phospho-c-Abl (panTyr) Sandwich ELISA Kit #7903能够检测内源性水平的Bcr-Abl或c-Abl蛋白，见图1.试剂盒的灵敏性见图2. 本试剂盒用来检测指示物种的蛋白，即通过内部测试确定的那些，也可以用来检测来自其他物种的同源蛋白。PathScan® Phospho-c-Abl (panTyr) Sandwich ELISA Kit是固相夹心酶联免疫吸附试验（ELISA），能够检测内源性水平的酪氨酸磷酸化Bcr-Abl和c-Abl蛋白。c-Abl兔抗包被微孔板。细胞裂解液孵育后，Bcr-Abl和c-Abl（磷酸和非磷酸）蛋白由包被的抗体捕获。充分冲洗后，加入磷酸酪氨酸鼠检测抗体检测捕获的酪氨酸磷酸化的Bcr-Abl和c-Abl蛋白。随后用抗鼠IgG，HRP-标记的抗体识别结合的检测蛋白。加入HRP底物TMB进行显色。显色的吸光度值与酪氨酸磷酸化的Bcr-Abl和c-Abl蛋白的量成正比。试剂盒中的抗体是针对试剂盒定制的。Bcr基因最初是通过嵌合Bcr-Abl癌基因鉴定出来的（1）。Bcr氨基末端区域含有一个寡聚化结构域、一个丝氨酸/苏氨酸激酶结构域和一个与SH2结构域结合的区域。蛋白质的中间有一个PH结构域和一个序列类似于GTP结合蛋白Rho家族鸟嘌呤核苷酸交换因子的区域。羧基末端区域可能被参与了小GTP结合蛋白Rac的GTP酶激活功能（2,3）。野生型细胞中Bcr的功能尚不清楚。PDGF受体可能使用Bcr作为下游信号的调解器（4）。研究显示，Bcr-Abl融合导致组成性激活的酪氨酸激酶产生，从而导致慢性粒细胞性白血病（CML）（5）。Bcr-Abl融合蛋白中Bcr酪氨酸177位点被磷酸化，这在改变Bcr-Abl活性中发挥了重要作用（6）。磷酸化的酪氨酸177位点为Gab2和GRB2提供了停靠位点（7,8）。

1. Wang, J.Y. et al. (2000) Oncogene 19, 5643-5650.
2. Van Etten, R.A. et al. (1999) Trends Cell. Biol. 9, 179-182.
3. Danial, N.N. et al. (2000) Oncogene 19, 2523-2531.
4. Shaul, Y. et al. (2000) Cell Death Differ. 7, 10-16.
5. Brasher, B.B. et al. (2000) J. Biol. Chem. 275, 35631-35637.
6. Pluk, H. et al. (2002) Cell 108, 247-259.

原厂资料：

Specificity / Sensitivity

PathScan® Phospho-c-Abl (panTyr) Sandwich ELISA Kit #7903 detects endogenous levels of Bcr-Abl or c-Abl protein when phosphorylated at Tyr residues in human cells, 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® Phospho-c-Abl (panTyr) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of tyrosine-phosphorylated Bcr-Abl and c-Abl proteins. A c-Abl rabbit antibody has been coated on the microwells. After incubation with cell lysates, Bcr-Abl and c-Abl protein (phospho and nonphospho) are captured by the coated antibody. Following extensive washing, a phospho-tyrosine mouse detection antibody is added to detect captured tyrosine-phosphorylated Bcr-Abl and c-Abl protein. Anti-mouse 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 tyrosine-phosphorylated Bcr-Abl and c-Abl protein.

Background

The Bcr gene was orginally identified by its presence in the chimeric Bcr-Abl oncogene (1). The amino-terminal region of Bcr contains an oligomerization domain, a serine/threonine kinase domain, and a region that binds SH2 domains. The middle of the protein has a PH domain and a region of sequence similarity to the guanine nucleotide exchange factors for the Rho family of GTP binding proteins. The carboxy-terminal region may be involved in a GTPase activating function for the small GTP-binding protein Rac (2,3). The function of wild type Bcr in cells remains unclear. PDGF receptor may use Bcr as a downstream signaling mediator (4). Research studies have shown that the Bcr-Abl fusion results in production of a constitutively active tyrosine kinase, which causes chronic myelogenous leukemia (CML) (5). Tyr177 of Bcr is phosphorylated in the Bcr-Abl fusion protein, which plays an important role in transforming the activity of Bcr-Abl (6). Phosphorylated Tyr177 provides a docking site for Gab2 and GRB2 (7,8).

1. Wang, J.Y. et al. (2000) Oncogene 19, 5643-5650.
2. Van Etten, R.A. et al. (1999) Trends Cell. Biol. 9, 179-182.
3. Danial, N.N. et al. (2000) Oncogene 19, 2523-2531.
4. Shaul, Y. et al. (2000) Cell Death Differ. 7, 10-16.
5. Brasher, B.B. et al. (2000) J. Biol. Chem. 275, 35631-35637.
6. Pluk, H. et al. (2002) Cell 108, 247-259.

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).