PathScan? Phospho-FLT3 (Tyr591) Chemiluminescent Sandwich ELISA Kit

• 产品编号：CST-7021C      品牌：CST       原厂货号：7021C
• 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：

PathScan® Phospho-FLT3 (Tyr591) Chemiluminescent Sandwich ELISA Kit #7021用于检测人细胞中内源磷酸化FLT3 (Tyr591)蛋白的表达。内部检测显示，本试剂盒能检测列举出的物种蛋白，但不排除用于其它物种同源蛋白检测的可能。PathScan® Phospho-FLT3 (Tyr591) Chemiluminescent Sandwich ELISA Kit利用固相双抗夹心酶联免疫检测法并结合化学发光读取系统，对内源性磷酸化的FLT3 (Tyr591)蛋白的表达进行检测。化学发光ELISA检测体系比传统的发色检测体现出更广的动态范围和更高的灵敏度。本化学发光ELISA检测在半量微孔板中进行，并在微量样本检测中呈现出更强的信号和检测灵敏度。微孔中已经包被了FLT3鼠单抗，与细胞裂解液共孵育后，磷酸化的和未磷酸化的FLT3均能被包被抗体捕获。多次洗板之后，加入磷酸化FLT3 (Tyr 591) 兔抗体对被捕获的磷酸化FLT3蛋白进行检测，然后加入抗兔IgG, HRP偶联抗体对检测抗体进行识别，最后加入化学发光检测试剂检测信号。按照相对光单位（RLU)，发射光强度与磷酸化FLT3 (Tyr591)蛋白的量成正比。FMS-相关的酪氨酸激酶3 (FLT3, 也称Flk2)是III型酪氨酸激酶受体家族的成员之一，其余成员包括c-Kit, PDGFR 和M-CSF 受体。FLT3表达在早期的造血前体细胞上，并支持造血系统的发育和分化（1,2）。FLT3结合其配体FL后激活，进而导致了酪氨酸酶自磷酸化和下游靶点酪氨酸磷酸化的级联反应（3）。FL刺激之后，FLT3就和PI3K激酶的p85亚基、 SHP2、 GRB2和Shc相关联（4-6）。Tyr589/591定位于FLT3的近膜区并对FLT3的酪氨酸激酶活性的调控有重要作用。20%的急性骨髓性白血病病人中发生FLT3内部串联重复 (ITDs)体细胞突变（7）。

1. Shurin, M.R. et al. (1998) Cytokine Growth Factor Rev 9, 37-48.
2. Naoe, T. et al. (2001) Cancer Chemother Pharmacol 48 Suppl 1, S27-30.
3. Namikawa, R. et al. (1996) Stem Cells 14, 388-95.
4. Beslu, N. et al. (1996) J Biol Chem 271, 20075-81.
5. Zhang, S. and Broxmeyer, H.E. (2000) Biochem Biophys Res Commun 277, 195-9.
6. Zhang, S. et al. (1999) J Leukoc Biol 65, 372-80.
7. Mizuki, M. et al. (2000) Blood 96, 3907-14.

原厂资料：

Specificity / Sensitivity

PathScan® Phospho-FLT3 (Tyr591) Chemiluminescent Sandwich ELISA Kit #7021 detects endogenous levels of phospho-FLT3 (Tyr591) in human cells. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Description

The PathScan® Phospho-FLT3 (Tyr591) Chemiluminescent Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of phospho-FLT3 (Tyr591) protein with a chemiluminescent readout. Chemiluminescent ELISAs often have a wider dynamic range and higher sensitivity than conventional chromogenic detection. This chemiluminescent ELISA, which is offered in low volume microplates, shows increased signal and sensitivity while using a smaller sample size. A FLT3 Mouse mAb has been coated on the microwells. After incubation with cell lysates, FLT3 (phospho or nonphospho) protein is captured by the coated antibody. Following extensive washing, a phospho-FLT3 (Tyr 591) Rabbit Antibody is added to detect the captured phospho-FLT3 protein. Anti-rabbit IgG, HRP-linked antibody is then used to recognize the bound detection antibody. Chemiluminescent reagent is added for signal development. The magnitude of light emission, measured in relative light units (RLU), is proportional to the quantity of phospho-FLT3 (Tyr591) protein.

Background

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).

1. Shurin, M.R. et al. (1998) Cytokine Growth Factor Rev 9, 37-48.
2. Naoe, T. et al. (2001) Cancer Chemother Pharmacol 48 Suppl 1, S27-30.
3. Namikawa, R. et al. (1996) Stem Cells 14, 388-95.
4. Beslu, N. et al. (1996) J Biol Chem 271, 20075-81.
5. Zhang, S. and Broxmeyer, H.E. (2000) Biochem Biophys Res Commun 277, 195-9.
6. Zhang, S. et al. (1999) J Leukoc Biol 65, 372-80.
7. Mizuki, M. et al. (2000) Blood 96, 3907-14.

注意事项：

Low volume microplate * 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).