A549 Cells GFP-STAT1
SIGMA/CLL1158
Product Type: Chemical
Fluorescence Microscopy A549 GFP-STAT1: Cat. No. CLL1158: A549 lung carcinoma knock-in cell line expressing the STAT1 gene (Signal Transducer and Activator of Transcription) endogenously tagged with GFP at the C-terminus. The cells were imaged live in Hank′s Balanced Salt Solution, Cat. No. H8264, supplemented with 2% Fetal Bovine Serum, Cat. No. F2442, 25 minutes after exposure to 100 ng/mL IFN-γ using GFP filterset (ex 450–490/em 500–550) and 40×/1.3 oil objective.
Differential Interference Contrast (DIC) Microscopy A549 GFP-STAT1: Cat. No. CLL1158: Differential interference contrast (DIC) microscopy of the same cells seen in the fluorescence microscopy image.
Fluorescence Microscopy A549 GFP-STAT1: Cat. No. CLL1158: Fluorescence image of the same cells taken before IFN-γ application.
| biological source | human male lung (Source Disease: Lung carcinoma) |
| OMIM accession no. | 600555   |
| 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: | This product is a human A549 cell line in which the genomic STAT1 gene has been endogenously tagged with a Green Fluorescent Protein (GFP) gene using CompoZr® zinc finger nuclease (ZFN) technology. Integration resulted in endogenous expression of the fusion protein in which GFP is attached to the C-terminus of STAT1. Fluorescence imaging shows STAT1-GFP expression throughout the cell. Upon activation with a ligand such as interferon-gamma (INF-γ), the cell line shows redistribution of STAT1 from the cytoplasm to the nucleus, making it useful for high content screening of compounds that modulate STAT1 activity. This stable cell line was expanded from a single clone. The target’s gene regulation and corresponding protein function are preserved in contrast to cell lines with overexpression via an exogenous promoter. |
| Application: | To learn more, please visit the Cellular Reporter 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: | Zinc Finger Nuclease (ZFN)-mediated targeted integration of a fluorescent GFP tag into the last coding exon just preceding the stop codon of the STAT1 gene on chromosome 2q32.2 to create a cell line exhibiting stable expression of the GFP transgene. The A549 cells are adherent, with a doubling time of approx. 22 hours. |
| General description: | A549 GFP-STAT1 are lung carcinoma cells from a human 58 year old caucasian male having two distinct ZFN modifications creating two fluorescent transgenes each expressed from their endogenous gene locus. This cell line was derived from ATCC Catalog No. CCL-185. |
| Legal Information: | CompoZr is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Preparation Note: | 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 in a 5% CO2 atmosphere. Subculture Ratio: approx. 1:3-1:8 The base medium for this cell line is F12 Ham Medium, Cat. No. N4888. To make the complete growth medium, add the following components to the base medium: fetal bovine serum, Cat. No. F4135, to a final concentration (v/v) of 10% and L-glutamine, Cat. No. G7513, at a final concentration of 2 mM. Cell freezing medium-DMSO 1X, Cat. No. C6164. |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 3 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | −196°C |
| UNSPSC | 41106514 |