Revision 1

#8951Store at +4C

 

()

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® EGF Receptor (L858R Mutant Specific) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of EGF receptor L858R mutant protein. An EGF receptor (L858R Mutant Specific) rabbit mAb has been coated on the microwells. After incubation with cell lysates, EGF receptor L858R mutant protein is captured by the coated antibody. Following extensive washing, an EGF receptor mouse monoclonal detection antibody is added to detect captured EGF receptor 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 EGF receptor L858R mutant protein.
Antibodies in kit are custom formulations specific to kit.

Specificity/Sensitivity

PathScan® EGF Receptor (L858R Mutant Specific) Sandwich ELISA Kit #8951 detects endogenous levels of EGF receptor L858R 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 trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    Limited Uses

    Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

    Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

    Revision 1
    #8951

    PathScan® EGF Receptor (L858R Mutant Specific) Sandwich ELISA Kit

    PathScan® EGF Receptor (L858R Mutant Specific) Sandwich ELISA Kit: Image 1 Expand Image
    Figure 1. EGFR mutant protein from H3255 cells can be detected by PathScan® EGF Receptor (L858R Mutant Specific) Sandwich ELISA Kit #8951. However, this kit cannot detect wild type EGFR protein from A-431 cells or EGFR E746-A750del protein from HCC827 cells. EGFR (either wild type or mutant) from these three cell lines can be detected by PathScan® Total EGF Receptor Sandwich ELISA Kit #7250, but no detection is found in MCF7 cells, which lack expression of EGFR. The absorbance readings at 450 nm are shown in the top figure, while the corresponding western blots using EGF Receptor (L858R Mutant Specific) (43B2) Rabbit mAb #3197, EGF Receptor (E746-A750del Specific) (6B6) XP® Rabbit mAb #2085, or EGF Receptor (C74B9) Rabbit mAb #2646 are shown in the bottom figure.
    PathScan® EGF Receptor (L858R Mutant Specific) Sandwich ELISA Kit: Image 2 Expand Image
    Figure 2. The relationship between protein concentration of lysates from A-431, H3255, or HCC827 cells and the absorbance at 450 nm is shown. A-431, H3255, or HCC827 cells (85% confluence) were harvested and then lysed.