A549 CELLS EGFR BIOSENSOR
SIGMA/CLL1097
Product Type: Chemical
Fluorescence Microscopy A549 CELLS EGFR BIOSENSOR: Cat. No. CLL1097: Lentiviral particles containing the EGFR Biosensor construct were used to transfect A549 lung carcinoma cells. Single cell clones were selected and assayed for activity with EGF. The clone in the image shows a homogenous response to translocation and internalization upon EGF stimulation. The cells were imaged live in Hanks′ Balanced Salt Solution, Cat. No. H8264, supplemented with 2% Fetal Bovine Serum, Cat. No. F2442, before EGF application using GFP filterset (ex 450–490/em 500–550) and 40×/1.3 oil objective.
Differential Interference Contrast (DIC) Microscopy A549 CELLS EGFR BIOSENSOR: Cat. No. CLL1097: Differential interference contrast (DIC) microscopy of the same cells seen in the fluorescence microscopy image.
Fluorescence Microscopy A549 CELLS EGFR BIOSENSOR: Cat. No. CLL1097: Fluorescence image of the same cells seen in the fluorescence microscopy image taken 4 minutes after addition of 100 ng/mL EGF.
Fluorescence Microscopy A549 CELLS EGFR BIOSENSOR: Cat. No. CLL1097: Florescence image of the same cells seen in the fluorescence microscopy image taken 45 minutes after addition of 100 ng/mL EGF.
| biological source | human male lung (Source Disease: Lung Carcinoma) |
| OMIM accession no. | 131550   |
| Quality Level | 200 |
| storage temp. | −196°C |
| Analysis Note: | 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. |
| Application: | 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. |
| Application: | To learn more, please visit the EGFR Biosensor Cell Line webpage |
| Disclaimer: | RESEARCH USE ONLY. This product is regulated in France when intended to be used for scientific purposes, including for import and export activities (Article L 1211-1 paragraph 2 of the Public Health Code). The purchaser (i.e. enduser) is required to obtain an import authorization from the France Ministry of Research referred in the Article L1245-5-1 II. of Public Health Code. By ordering this product, you are confirming that you have obtained the proper import authorization. |
| Features and Benefits: | Lentiviral transduction of DNA construct for stable expression of GFP-tagged SH2 binding domain. |
| Features and Benefits: | These A549 cells are adherent with a doubling time of approx. 22 hours. |
| General description: | This cell line was derived from ATCC catalog No. CCL-185. |
| Preparation Note: | Cell freezing medium-DMSO 1X, Cat. No. C6164 |
| Preparation Note: | Media renewal changes two to three times per week. |
| Preparation Note: | 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. |
| Preparation Note: | 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. |
| WGK Germany | WGK 3 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | −196°C |
| UNSPSC | 41106514 |