Cell Signaling Technology

Product Pathways - Tyrosine Kinase / Adaptors

PathScan® Phospho-Ret (panTyr) Chemiluminescent Sandwich ELISA Kit #12321

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No. Size Price
12321C 1 Kit ( 96 assays ) ¥6,345.00 现货查询 购买询价
Product Includes Volume Solution Color
ELISA Sample Diluent 25 ml Blue
Phospho Tyrosine Mouse Detection mAb 1 ea Green (Lyophilized)
Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated) 1 ea Red (Lyophilized)
Detection Antibody Diluent 5.5 ml Green
HRP Diluent 5.5 ml Red
Luminol/Enhancer Solution 3 ml Colorless
ELISA Wash Buffer (20X) 25 ml Colorless
Cell Lysis Buffer (10X) #9803 15 ml Yellowish
Sealing Tape 2 sheets
Stable Peroxide Buffer 3 ml Colorless
Ret Rabbit mAb Coated Microwells 96 tests

Specificity / Sensitivity

PathScan® Phospho-Ret (panTyr) Chemiluminescent Sandwich ELISA Kit #12321 detects endogenous levels of tyrosine-phosphorylated Ret protein 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.

PathScan® Phospho-Ret (panTyr) Chemiluminescent Sandwich ELISA Kit #12321能够检测人细胞中内源性水平的酪氨酸磷酸化的Ret蛋白。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。

Description

The PathScan® Phospho-Ret (panTyr) Chemiluminescent Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of tyrosine-phosphorylated Ret 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 Ret Rabbit mAb has been coated onto the microwells. After incubation with cell lysates, Ret protein (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a Phospho-Tyrosine Mouse Detection mAb is added to detect captured tyrosine-phosphorylated Ret protein. Anti-mouse 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 tyrosine-phosphorylated Ret protein. 
 Antibodies in kit are custom formulations specific to kit.

PathScan® Phospho-Ret (panTyr) Chemiluminescent Sandwich ELISA Kit是固相夹心酶联免疫吸附试验(ELISA),用化学发光法检测内源性水平的酪氨酸磷酸化Ret蛋白。化学发光酶联免疫吸附比传统的显色检测往往有更广阔的动态范围和更高的灵敏度。此化学发光ELISA提供在低量微孔板,提高信号灵敏度,同时使用一个较小的样本量。微孔包被Ret兔单克隆抗体。与细胞裂解物孵育后,Ret蛋白(磷酸化和非磷酸化)由包被的抗体捕获。充分洗涤后,加入磷酸化酪氨酸小鼠检测单克隆抗体,以检测捕获得酪氨酸磷酸化Ret蛋白。然后,使用抗小鼠IgG,HRP-标记的抗体,识别结合的检测抗体。加入化学发光试剂进行显色。发射光强度用相对光单位(RLU)测量,并与总EGF受体蛋白的量成正比。试剂盒中的抗体是针对试剂盒特别定制的。

Figure 1. The relationship between protein concentration of lysates, untreated (Nonphospho) and phosphatase inhibitor treated (Phospho), and immediate light generation with chemiluminescent substrate is shown. TT cells were cultured (85% confluent) and lysed with the addition of phosphatase inhibitor to the PathScan® Sandwich ELISA Cell Lysis Buffer (1X) #9803. Graph inset corresponding to the shaded area shows high sensitivity and a linear response at the low protein concentration range.图1.如图所示裂解物蛋白浓度之间的关系,细胞不处理(非磷酸化)或用磷酸酶抑制剂处理(磷酸化),立即产生光化学发光底物。TT细胞培养(85%汇合)后,加入磷酸酶抑制剂,用PathScan® Sandwich ELISA Cell Lysis Buffer (1X) #9803进行裂解。图中对应的阴影部分的面积图形显示了高灵敏度和低蛋白质浓度范围内的线性响应。

Background

The Ret proto-oncogene (c-Ret) is a receptor tyrosine kinase that functions as a multicompetent receptor complex in conjunction with other membrane-bound, ligand-binding GDNF family receptors (1). Ligands that bind the Ret receptor include the glial cell line-derived neurotropic factor (GDNF) and its congeners neurturin, persephin, and artemin (2-4). Research studies have shown that alterations in the corresponding RET gene are associated with diseases including papillary thyroid carcinoma, multiple endocrine neoplasia (type 2A and 2B), familial medullary thyroid carcinoma, and a congenital developmental disorder known as Hirschsprung’s disease (1,3). The Tyr905 residue located in the Ret kinase domain plays a crucial role in Ret catalytic and biological activity. Substitution of Phe for Tyr at position 905 dramatically inhibits Ret autophosphorylation activity (5).

Ret原癌基因(c-Ret)是一个多功能的酪氨酸激酶受体复合物,它和其它膜结合的GDNF家族的配体结合型受体共同发挥作用(1)。与Ret受体结合的配体包括神经胶质细胞来源的神经营养因子(GDNF)和它的同类物neurturin、 persephin 和 artemin (2-4)。在相应的Ret 基因的改变会引起乳突状甲状腺癌、多发性内分泌肿瘤(2A和2B型),家族性髓样甲状腺癌和先天性巨结肠病(1,3)。位于Ret激酶区的Tyr905位点对于Ret的催化功能和生物学功能十分重要。替换Tyr905的Phe能显著抑制Ret的自磷酸化活性(5)。

  1. Airaksinen, M.S. et al. (1999) Mol Cell Neurosci 13, 313-25.
  2. Takahashi, M. et al. (1989) Oncogene 4, 805-6.
  3. Manié, S. et al. (2001) Trends Genet 17, 580-9.
  4. Tallini, G. and Asa, S.L. (2001) Adv Anat Pathol 8, 345-54.
  5. Iwashita, T. et al. (1999) Oncogene 18, 3919-22.

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For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

PathScan is a trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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