| 货号 | 7245C |
| 描述 | PathScan® Total Notch1 Sandwich ELISA Kit from Cell Signaling Technology is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Notch1. 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 Notch1 rabbit 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 Notch1. |
| 反应种属 | Human |
| 应用 | ELISA |
| 目标/特异性 | PathScan® Total Notch1 Sandwich ELISA Kit #7245 detects endogenous levels of Notch1 protein 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. |
| 供应商 | CST |
| 背景 | 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. |
| 存放说明 | 4C and -20C |
| 参考文献 | Artavanis-Tsakonas, S. et al. (1999) Science 284, 770-6. Chan, Y.M. and Jan, Y.N. (1998) Cell 94, 423-6. Schroeter, E.H. et al. (1998) Nature 393, 382-6. Rand, M.D. et al. (2000) Mol Cell Biol 20, 1825-35. |
Figure 2. The relationship between the protein concentration of lysates from Jurkat cells, untreated or treated with γ secretase inhibitor, and the absorbance at 450 nm using the PathScan® Total Notch1 Sandwich ELISA Kit #7245 is shown. 图2，使用PathScan® Total Notch1 Sandwich ELISA Kit #7245检测未处理或经过 γ 分泌酶处理的Jurkat细胞裂解物，蛋白浓度与450 nm处吸光值间关系如图所示。 | |
Figure 1. Both cleaved and noncleaved Notch1 can be detected by the PathScan® Total Notch1 Sandwich ELISA Kit #7245. Jurkat cells express cleaved Notch1 (Val1744) and γ secretase inhibitor inhibits the cleavage of Notch1. Jurkat cells were treated with 25 μM γ secretase inhibitor for 96 hours. The absorbance readings at 450 nm are shown on the top, while the corresponding western blots using Notch1 (D1E11) XP® Rabbit mAb #3608 (left panel) or Cleaved Notch1 (Val1744) Antibody #2421 (right panel) are shown on the bottom. 图1.PathScan® Total Notch1 Sandwich ELISA Kit #7245可以检测剪切后和未剪切的Notch1蛋白.Jurkat细胞表达剪切后的Notch1，γ 分泌酶抑制剂可以抑制这种剪切。用25μM γ 分泌酶抑制剂处理Jurkat细胞96小时。450nm处吸光值如图上部所示，使用Notch1 (D1E11) XP® Rabbit mAb兔单抗#3608 (左)或Cleaved Notch1 (Val1744)抗体#2421（右）进行的western blot实验如图下部所示。 |
