Revision 5

#7890Store at +4C

1 个试剂盒

(96 assays)

Species Cross Reactivity

H

UniProt ID:

#P21860

Entrez-Gene Id:

#2065

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Color Storage Temp
HER3/ErbB3 Rabbit mAb Coated Microwells 97125 96 tests +4C
Phospho Tyrosine Mouse Detection mAb 12982 1 ea Green (Lyophilized) +4C
Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated) 13304 1 ea Red (Lyophilized) +4C
Detection Antibody Diluent 13339 11 ml Green +4C
HRP Diluent 13515 11 ml Red +4C
TMB Substrate 7004 11 ml +4C
STOP Solution 7002 11 ml +4C
Sealing Tape 54503 2 ea +4C
ELISA Wash Buffer (20X) 9801 25 ml +4C
ELISA Sample Diluent 11083 25 ml Blue +4C
Cell Lysis Buffer (10X) 9803 15 ml -20C

*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.

Description

The PathScan® Phospho-HER3/ErbB3 (panTyr) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of tyrosine-phosphorylated HER3/ErbB3 protein. A HER3/ErbB3 rabbit mAb has been coated on the microwells. After incubation with cell lysates, HER3/ErbB3 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 HER3/ErbB3 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 HER3/ErbB3 protein phosphorylated on tyrosine.

*Antibodies in this kit are custom formulations specific to kit.

Specificity/Sensitivity

PathScan® Phospho-HER3/ErbB3 (panTyr) Sandwich ELISA Kit #7890 detects endogenous levels of HER3/ErbB3 protein only when phosphorylated at Tyr residues (see Figure 1). The kit sensitivity is shown in Figure 2. This kit does not cross-react with other proteins of ErbB family (data not shown). This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Background

HER3/ErbB3 is a member of the ErbB receptor protein tyrosine kinase family, but it lacks tyrosine kinase activity. Tyrosine phosphorylation of ErbB3 depends on its association with other ErbB tyrosine kinases. Upon ligand binding, heterodimers form between ErbB3 and other ErbB proteins, and ErbB3 is phosphorylated on tyrosine residues by the activated ErbB kinase (1,2). There are at least 9 potential tyrosine phosphorylation sites in the carboxy-terminal tail of ErbB3. These sites serve as consensus binding sites for signal transducing proteins, including Src family members, Grb2, and the p85 subunit of PI3 kinase, which mediate ErbB downstream signaling (3). Both Tyr1222 and Tyr1289 of ErbB3 reside within a YXXM motif and participate in signaling to PI3K (4).

Investigators have found that ErbB3 is highly expressed in many cancer cells (5) and activation of the ErbB3/PI3K pathway is correlated with malignant phenotypes of adenocarcinomas (6). Research studies have demonstrated that in tumor development, ErbB3 may function as an oncogenic unit together with other ErbB members (e.g., ErbB2 requires ErbB3 to drive breast tumor cell proliferation) (7). Thus, investigators view inhibiting interaction between ErbB3 and ErbB tyrosine kinases as a novel strategy for anti-tumor therapy.

  1. Yarden, Y. and Sliwkowski, M.X. (2001) Nat Rev Mol Cell Biol 2, 127-37.
  2. Guy, P.M. et al. (1994) Proc Natl Acad Sci U S A 91, 8132-6.
  3. Songyang, Z. et al. (1993) Cell 72, 767-78.
  4. Kim, H.H. et al. (1994) J Biol Chem 269, 24747-55.
  5. Sithanandam, G. et al. (2003) Carcinogenesis 24, 1581-92.
  6. Kobayashi, M. et al. (2003) Oncogene 22, 1294-301.
  7. Holbro, T. et al. (2003) Proc Natl Acad Sci U S A 100, 8933-8.

Background References

    Cross-Reactivity Key

    H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    PathScan is a registered trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    限制使用

    除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

    专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

    Revision 5
    #7890

    PathScan® Phospho-HER3/ErbB3 (panTyr) Sandwich ELISA Kit

    PathScan® Phospho-HER3/ErbB3 (panTyr) Sandwich ELISA Kit: Image 1 Expand Image
    图 1. 使用 PathScan® Phospho-HER3/ErbB3 (panTyr) Sandwich ELISA Kit #7890(右上图)可检测在磷酸酶抑制剂*中裂解的 Calu-3 细胞(磷酸裂解物)中 HER3/ErbB3 的组成性磷酸化。相反,在未经磷酸酶抑制剂*裂解的 Calu-3 细胞(非磷酸裂解物)中则检测到低水平的磷酸-HER3/ErbB3 蛋白。使用 PathScan® Total HER3/ErbB3 Sandwich ELISA Kit #7888(左上图)可在非磷酸或磷酸裂解物中检测到类似水平的 HER3/ErbB3 总蛋白。在 450 nm 处的吸光度如顶部图所示,使用 Phospho-HER3/ErbB3 (Tyr1289) Rabbit mAb #4791(右图)或 total HER3/ErbB3 Antibody #4754(左图)检测到的相应蛋白质印迹如底部图所示。*磷酸酶抑制剂包括焦磷酸钠、β-甘油磷酸和 Na3VO4
    PathScan® Phospho-HER3/ErbB3 (panTyr) Sandwich ELISA Kit: Image 2 Expand Image
    图 2. 未经处理和已经 NRG 处理的 T-47D 细胞的裂解物蛋白浓度与使用试剂盒测得的光密度读数之间的关系。饥饿后,T-47D 细胞(85% 融合度)在 37°C 下用 Human Neuregulin-1 (hNRG-1) #5218 (100 ng/ml) 处理 5 分钟后进行裂解。