Cell Signaling Technology

Product Pathways - Tyrosine Kinase / Adaptors

EGF Receptor (L858R Mutant Specific) (43B2) Rabbit mAb #3197

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IHC-P IF-P IF-IC F H Endogenous 175 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-P=Immunofluorescence (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

* Product-specific protocol.

Specificity / Sensitivity

EGFR (L858R Mutant Specific) (43B2) Rabbit mAb detects endogenous levels of EGFR mutant L858R protein. Careful titration of this antibody may be required to obtain optimal specificity.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to L858R mutant sequence of human EGF receptor.

Western Blotting

Western Blotting

Western blot analysis of extracts from various EGFR expressing cell lines, showing the specificity of detection of mutant EGFR (L858R) protein using EGF Receptor (L858R Mutant Specific) (43B2) Rabbit mAb (upper), and total EGFR control antibody (lower).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded H1975 (left, EGFR L858R positive) and H1650 xenograft (right, EGFR deletion positive) using EGF Receptor (L858R Mutant Specific) (43B2) Rabbit mAb (upper) and total EGFR (D38B1) XP® Rabbit mAb #4267 (lower).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma of known mutational status using EGF Receptor (L858R Mutant Specific) (43B2) Rabbit mAb (upper) and total EGFR (D38B1) XP® Rabbit mAb #4267 (lower). EGFR L858R positive lung (left), EGFR deletion positive lung sections (right).


Flow Cytometry

Flow Cytometry

Flow cytometric analysis of two non-small cell lung cancer cell lines using EGF Receptor (L858R Mutant Specific) (43B2) Rabbit mAb. H3255 cells (green) harbor the L858R EGFR mutation while the H1650 cells (blue) have the exon 19 mutation (delE746-A750).

IF-IC

IF-IC

Confocal immunofluorescent analysis of H3255 (left), HCC827 (middle) and A431 cells (right) using EGF Receptor (L858R Mutant Specific) (43B2) Rabbit mAb (green) and MEK1/2 (L38C12) Mouse mAb #4694 (red). H3255 cells have the L858R EGFR mutation, HCC827 cells have an exon 19 EGFR deletion, and A431 cells overexpress wild type EGFR. Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

IF-P

IF-P

Confocal immunofluorescent analysis of paraffin-embedded H3255, H1975, HCC827 and H1650 xenografts using EGF Receptor (L858R Mutant Specific) (43B2) Rabbit mAb (upper, green) and EGFR (D38B1) XP® Rabbit mAb #4267 (lower, green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).


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 adenocarinomas that respond to EGFR inhibitors, such as gefinitib 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.

Application References

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Companion Products

The manufacture, use, sale and import of this product is within the scope of one or more intellectual property rights (including patents and patent applications) owned or controlled by Cell Signaling Technology. The purchase of this product conveys to the buyer a non-transferrable right to use the purchased product only in research conducted by the buyer. The sale of the product is expressly conditioned on the buyer not using the products or its components (1) to analyze or reverse engineer the product for its chemical/physical properties and composition (including e.g., identification of the sequence); (2) in manufacturing; (3) to provide a service, information, or data to an unaffiliated third party for payment; (4) for therapeutic, diagnostic or prophylactic purposes; (5) resale, whether or not such product are resold for use in research; or for any other commercial purpose. For information on purchasing a license to this product for purposes other than research, contact Cell Signaling Technology, Inc. Business Development at busdev@cellsignal.com. Rabbit monoclonal antibody produced under license (granting certain rights including those under U. S. Patents No. 5,675,063 and 7,429,487) from Epitomics, Inc.

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

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