| 货号 | 7761C |
| 描述 | CSTs PathScan® Phospho-FLT3 (panTyr) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of tyrosine-phosphorylated FLT3 protein. A FLT3 rabbit antibody has been coated on the microwells. After incubation with cell lysates, FLT3 protein (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a phospho-tyrosine mouse detection antibody is added to detect captured tyrosine-phosphorylated FLT3 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 FLT3 protein phosphorylated on tyrosine. |
| 反应种属 | Human |
| 应用 | ELISA |
| 目标/特异性 | PathScan® Phospho-FLT3 (panTyr) Sandwich ELISA Kit #7761 detects endogenous levels of FLT3 protein 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. |
| 供应商 | CST |
| 背景 | FMS-related tyrosine kinase 3 (FLT3, also called Flk2), is a member of the type III receptor tyrosine kinase family, which includes c-Kit, PDGFR and M-CSF receptors. FLT3 is expressed on early hematopoietic progenitor cells and supports growth and differentiation within the hematopoietic system (1,2). FLT3 is activated after binding with its ligand FL, which results in a cascade of tyrosine autophosphorylation and tyrosine phosphorylation of downstream targets (3). The p85 subunit of PI3 kinase, SHP2, GRB2 and Shc are associated with FLT3 after FL stimulation (4-6). Tyr589/591 is located in the juxtamembrane region of FLT3 and may play an important role in regulation of FLT3 tyrosine kinase activity. Somatic mutations of FLT3 consisting of internal tandem duplications (ITDs) occur in 20% of patients with acute myeloid leukemia (7). |
| 存放说明 | 4C |
| 参考文献 | Shurin, M.R. et al. (1998) Cytokine Growth Factor Rev 9, 37-48. Naoe, T. et al. (2001) Cancer Chemother Pharmacol 48 Suppl 1, S27-30. Namikawa, R. et al. (1996) Stem Cells 14, 388-95. Beslu, N. et al. (1996) J Biol Chem 271, 20075-81. Zhang, S. and Broxmeyer, H.E. (2000) Biochem Biophys Res Commun 277, 195-9. Zhang, S. et al. (1999) J Leukoc Biol 65, 372-80. Mizuki, M. et al. (2000) Blood 96, 3907-14. |
Figure 1: Constitutive phosphorylated of FLT3 in SEM cells is detected by PathScan® Phospho-FLT3 (panTyr) Sandwich ELISA kit #7761. In contrast, only a low level of phospho-FLT3 protein is detected in SEM cells treated with CT-53518, an inhibitor of FLT3 tyrosine phosphorylation. The inhibitor does not affect the level of total FLT3 protein detected by PathScan® Total FLT3 Sandwich ELISA kit #7202. Absorbance at 450 nm is shown in the top figure, while the corresponding western blots using Phospho-FLT3 (Tyr591) Antibody #3461 (right panel) or FLT3 Rabbit mAb #3462 (left panel), is shown in the bottom figure.图1：用PathScan® Phospho-FLT3 (panTyr) Sandwich ELISA kit #7761能够检测到磷酸化的FLT3在SEM细胞上的组成型表达。对照，在经过FLT3酪氨酸磷酸酶抑制剂CT-53518处理过的SEM细胞裂解液中仅能检测到低水平的phospho-FLT3存在。该抑制剂不影响PathScan® Total FLT3 Sandwich ELISA kit #7202对总FLT3表达水平的检测。上图显示在波长450nm处的光吸收值，下图显示用Phospho-FLT3 (Tyr591) Antibody #3461（右）或FLT3 Rabbit mAb #3462（左）进行免疫印迹检测的结果。 | |
Figure 2: The relationship between total protein concentration of the lysate from untreated or inhibitor-treated SEM cells and absorbance at 450 nm is shown. Unstarved SEM cells (10-6 cells/ml) were left untreated or treated with CT-53518 (50 ng/ml) for 120 min and then lysed.图2：未处理的或抑制剂处理过的SEM细胞的裂解液中蛋白含量和450nm吸光值之间的关系。未饥饿的SEM细胞(106 cells/ml)不经或经过CT-53518 (50 ng/ml) 处理 120 分钟后进行裂解。 |
