The EGFR Biosensor cell line consists of the SH2 domain of the adaptor protein Grb2 fused to GFP. The biosensor was transduced into the A549 cell line and a stable cell line was cloned from a single cell. Within minutes after stimulation of the cell line with EGF, the biosensor shows robust redistribution towards the cell membrane and subsequent internalization through endocytosis. The internalization kinetics of the sensor-receptor complex can be quantified by image analysis software to count granules. The reporter response is highly selective as it can be induced only by specific EGFR ligands and is abolished by a selective inhibitor of EGFR, Tyrphostin AG 1478.
To learn more, please visit the EGFR Biosensor Cell Line webpage
Lentiviral transduction of DNA construct for stable expression of GFP-tagged SH2 binding domain.
These A549 cells are adherent with a doubling time of approx. 22 hours.
Tested for Mycoplasma, sterility, post-freeze viability, short terminal repeat (STR) analysis for cell line identification, cytochrome oxidase I (COI) analysis for cell line species confirmation.
Cell freezing medium-DMSO 1X, Cat. No. C6164
Media renewal changes two to three times per week.
Rapidly thaw vial by gentle agitation in 37°C water bath (~2 minutes), keeping vial cap out of the water. Decontaminate with 70% ethanol, add 9 mL culture media and centrifuge 125 x g (5-7 minutes). Resuspend in complete culture media and incubate at 37°C.
The base medium for this cell line is RPMI-1640, Cat. No. R0883. To make the complete growth medium, add the following components to the base medium: fetal bovine serum, Cat. No. F2442, to a final concentration (v/v) of 10%, L-glutamine, Cat. No. G7513, at a final concentration of 2 mM and puromycin, Cat. No. P9620, at a final concentration of 1 μg/mL.