Notch Isoform Antibody Sampler Kit

• 产品编号：CST-3640S      品牌：Cell Signaling Technology       原厂货号：3640S
• 产品分类：抗体 > 一抗 > 复合抗体试剂盒
• 应用分类：

描述：

Cleaved Notch1 (Val1744) (D3B8) XP^® Rabbit mAb兔单抗只识别1743位甘氨酸和1744位缬氨酸之间被剪切的内源性Notch1蛋白的胞质结构域。抗体不识别全长Notch1蛋白或者被其他位点的剪切型Notch1。在不同细胞中存在Notch1的远处羧基末端的突变，因此此抗体识别大小不同的剪切型Notch1（见参考文献6）Notch1 (D6F11) XP^® Rabbit mAb兔源抗体检测内源性的总Notch1蛋白。它识别全长(~300 kDa)和跨膜/胞内区域NTM(~120 kDa)。Notch2 (D67C8) XPTM兔源单抗识别内源性的Notch2蛋白。它识别全长(~300 kDa)和跨膜/胞内区域NTM(~110 kDa)。Notch3 (D11B8) Rabbit mAb兔抗体识别内源性的总Notch3蛋白。此抗体识别全长270kDa的Notch3和截短的蛋白(NTM)，包含短的胞外区域，跨膜区域和胞内区域（90kDa）。各种单抗由合成肽段免疫动物产生，相应肽段分别与人源Notch1蛋白邻近1744位缬氨酸剪切位点的氨基酸序列一致，如人类Notch1的2497位甘氨酸，人类Notch2的2332位缬氨酸或人类Notch3的2312位甘氨酸。Notch Isoform Antibody Sampler Kit提供经济合适的方式检测Notch信号通路。试剂盒含有足够做四次小的Western blot所需的一抗和二抗。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。组成型激活的Notch1信号通路与大部分T细胞急性淋巴母细胞白血病（T-ALL）有关。Notch1信号通路的活化与Notch1自身突变或泛素连接酶组件FBW7有关（5-6）。Notch2是Notch家族成员，Notch2突变与Alagille综合征有关（7）。Notch3是Notch家族成员，加工过程与Notch1相似（1）。它主要表达在动脉平滑肌细胞（SMC）。在Notch3的胞外区域突变改变了半胱氨酸残基数量，这与大脑常染色体显性动脉病皮质下梗死和脑白质病（CADASIL）有关, （CADASIL）为一种导致成人中风痴呆的遗传血管病（9-11）。近来研究显示Notch3在多种癌症过表达（12-14）。

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.
5. Weng, A.P. et al. (2004) Science 306, 269-71.
6. Thompson, B.J. et al. (2007) J Exp Med 204, 1825-35.
7. McDaniell, R. et al. (2006) Am J Hum Genet 79, 169-73.
8. Baron, M. (2003) Semin Cell Dev Biol 14, 113-9.
9. Kalimo, H. et al. (2002) Brain Pathol 12, 371-84.
10. Karlström, H. et al. (2002) Proc Natl Acad Sci U S A 99, 17119-24.
11. Monet, M. et al. (2007) Hum Mol Genet 16, 982-92.
12. Park, J.T. et al. (2006) Cancer Res 66, 6312-8.
13. Gramantieri, L. et al. (2007) Liver Int 27, 997-1007.
14. Yamaguchi, N. et al. (2008) Cancer Res 68, 1881-8.

原厂资料：

Specificity / Sensitivity

Cleaved Notch1 (Val1744) (D3B8) XP^® Rabbit mAb detects endogenous levels of the cytosolic domain of Notch1 only when cleaved between Gly1743 and Val1744. The antibody does not recognize full length Notch1 or Notch1 cleaved at other positions. It detects cleaved Notch1 with different sizes in different cell lines due to mutations at the distant C-termini of Notch1 (see ref 6). Notch1 (D6F11) XP^® Rabbit detects endogenous level of total notch1 protein. It recognizes both the full-length (~300 kDa) and the transmembrane/intracellular region NTM (~120 kDa). Notch2 (D67C8) XPTM Rabbit mAb detects endogenous levels of total Notch2 protein. It recognizes both the full-length (~ 300 kDa) and the transmembrane/intracellular region NTM (~110 kDa). Notch3 (D11B8) Rabbit mAb detects endogenous levels of total Notch3 protein. The antibody recognizes both full-length (FL) Notch3 at 270 kDa and a truncated protein (NTM) containing a short extracellullar region, the transmembrane domain and the intracellular region at 90 kDa.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the sequence at the Val1744 cleavage site of human Notch1, Gln2497 of human Notch1, Val2332 of human Notch2, or Glu2312 of human Notch3 protein.

Description

The Notch Isoform Antibody Sampler Kit provides an economical means to investigate Notch Signaling. The kit contains primary and secondary antibodies to perform four western mini-blots with each antibody.

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.

Constitutively activated Notch1 signaling is associated with the majority of cases of T cell acute lymphoblastic leukemia (T-ALL). The activation is either due to mutations in Notch1 itself or in the components of ubiquitin ligase complex, namely FBW7 (5-6). Notch2 is a member of the Notch family and mutation in Notch2 is associated with Alagille syndrome (7). Notch3 is a member of the Notch family and is processed similar to Notch1 (8). It is expressed primarily in arterial smooth muscle cells (SMC). Mutations altering the number of cysteine residues in the notch3 extracellular region are associated with cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL), a hereditary angiopathy leading to strokes and dementia in adults (9-11). Recent studies indicate that Notch3 is overexpressed in many types of cancer (12-14).

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.
5. Weng, A.P. et al. (2004) Science 306, 269-71.
6. Thompson, B.J. et al. (2007) J Exp Med 204, 1825-35.
7. McDaniell, R. et al. (2006) Am J Hum Genet 79, 169-73.
8. Baron, M. (2003) Semin Cell Dev Biol 14, 113-9.
9. Kalimo, H. et al. (2002) Brain Pathol 12, 371-84.
10. Karlström, H. et al. (2002) Proc Natl Acad Sci U S A 99, 17119-24.
11. Monet, M. et al. (2007) Hum Mol Genet 16, 982-92.
12. Park, J.T. et al. (2006) Cancer Res 66, 6312-8.
13. Gramantieri, L. et al. (2007) Liver Int 27, 997-1007.
14. Yamaguchi, N. et al. (2008) Cancer Res 68, 1881-8.

注意事项：

Storage: Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM
NaCl, 100 µg/ml BSA and 50% glycerol and less than 0.02%
sodium azide. Store at –20°C. Do not aliquot the antibodies.