Figure 1. EGFR mutant protein from HCC827 cells can be detected by PathScan® EGF Receptor (E746-A750del Specific) Sandwich ELISA Kit #8950. However, this kit cannot detect wild type EGFR protein from A-431 cells or EGFR (L858R) mutant protein from H3255 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) Rabbit mAb #2085, or EGF Receptor (C74B9) Rabbit mAb #2646 are shown in the bottom figure.
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.
The PathScan® EGF Receptor (E746-A750del Specific) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of EGF receptor E746-A750del mutant protein. An EGF receptor (E746-A750del Specific) rabbit mAb 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, 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 E746-A750del mutant protein.
Antibodies in kit are custom formulations specific to kit.
PathScan® EGF Receptor (E746-A750del Specific) Sandwich ELISA Kit #8950 detects endogenous levels of EGF receptor 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.
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.
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