Revision 1

#7702Store at +4C

 

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Species Cross Reactivity

H

UniProt ID:

#P00533

Entrez-Gene Id:

#1956

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
TMB Substrate 7004 11 ml Colorless +4C
STOP Solution 7002 11 ml Colorless +4C
Sealing Tape 54503 2 ea +4C
ELISA Wash Buffer (20X) 9801 25 ml Colorless +4C
ELISA Sample Diluent 11083 25 ml Blue +4C
Cell Lysis Buffer (10X) 9803 15 ml Yellowish -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-EGF Receptor (Tyr1068) (E746-A750del Specific) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Phospho-EGF receptor (Tyr1068) (E746-A750del) mutant protein. An EGF Receptor (E746-A750del Specific) Rabbit Antibody has been coated on the microwells. After incubation with cell lysates, EGF receptor (E746-A750del) mutant protein (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a Phospho-EGF Receptor (Tyr1068) Mouse Detection Antibody is added to detect captured phosphorylated EGF receptor (E746-A750del) mutant 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 phospho-EGF receptor (Tyr1068) (E746-A750del) mutant protein.
Antibodies in kit are custom formulations specific to kit.

Specificity/Sensitivity

PathScan® Phospho-EGF Receptor (Tyr1068) (E746-A750del Specific) Sandwich ELISA Kit #7702 detects endogenous levels of phospho-EGF receptor (Tyr1068) (E746-A750del) mutant protein 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.

Background

The epidermal growth factor (EGF) receptor is a 170 kDa transmembrane tyrosine kinase that belongs to the HER/ErbB protein family. Research studies have shown that somatic mutations in the tyrosine kinase domain of EGF receptor (EGFR) are present in a subset of lung adenocarcinomas that respond to EGFR inhibitors, such as gefitinib and erlotinib (1-3). Two types of mutations account for approximately 90% of mutated cases: a specific point mutation, L858R, that occurs in exon 21 and short in-frame deletions in exon 19 (4,5). The most frequent exon 19 deletion is E746-A750, accounting for 90% of lesions at this site, although some rare variants occur.

  1. Lynch, T.J. et al. (2004) N Engl J Med 350, 2129-39.
  2. Pao, W. et al. (2004) Proc Natl Acad Sci USA 101, 13306-11.
  3. Haber, D.A. et al. (2005) Cold Spring Harb Symp Quant Biol 70, 419-26.
  4. Kosaka, T. et al. (2004) Cancer Res 64, 8919-23.
  5. Riely, G.J. et al. (2006) Clin Cancer Res 12, 7232-41.

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 1
    #7702

    PathScan® Phospho-EGF Receptor (Tyr1068) (E746-A750del Specific) Sandwich ELISA Kit

    PathScan® Phospho-EGF Receptor (Tyr1068) (E746-A750del Specific) Sandwich ELISA Kit: Image 1 Expand Image
    图 1. 使用 PathScan® Phospho-EGF Receptor (Tyr1068) (E746-A750del Specific) Sandwich ELISA Kit #7702 在添加磷酸酶抑制剂*裂解的 HCC827 细胞(磷酸化裂解物)中检测到 EGF 受体 (E746-A750del) 突变蛋白的组成性磷酸化。相比之下,在未添加磷酸酶抑制剂*裂解的 HCC827 细胞(非磷酸化裂解物)中,检测到低水平的磷酸化 EGF 受体 (Tyr1068) (E746-A750del) 蛋白。该试剂盒不能检测 A-431 细胞中的野生型 EGFR 蛋白(磷酸化或非磷酸化形式),也不可检测 H3255 细胞中的 EGFR (L858R) 突变蛋白(磷酸化或非磷酸化形式)。在 450 nm 处的吸光度如顶部图所示(仅在 HCC827 磷酸化裂解物中可见阳性信号),使用 Phospho-EGF Receptor (Tyr1068) (D7A5) XP® Rabbit mAb #3777 检测到的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-EGF Receptor (Tyr1068) (E746-A750del Specific) Sandwich ELISA Kit: Image 2 Expand Image
    图 2. 磷酸或非磷酸裂解物的蛋白浓度与在 450 nm 处的吸光度之间的关系如图所示。HCC827 细胞培养(85% 融合度)后,在添加或未添加磷酸酶抑制剂的裂解缓冲液中进行裂解(分别为磷酸化裂解物或非磷酸化裂解物)。