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CLL1140

Sigma-Aldrich

A549 Cells RFP-STAT3

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NACRES:
NA.81

biological source

human male lung (Source Disease: Lung carcinoma)

Quality Level

OMIM accession no.

storage temp.

−196°C

Gene Information

human ... STAT3(6774)

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CLL1140

A549 Cells RFP-STAT3

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A549 Cells GFP-STAT1

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CLL1167

A549 Cells GFP-SMAD4

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CLL1149

A549 Cells GFP-CTNNB1 RFP-LMNB1

Quality Level

200

Quality Level

200

Quality Level

200

Quality Level

200

Gene Information

human ... STAT3(6774)

Gene Information

human ... STAT1(6772)

Gene Information

human ... SMAD4(4089)

Gene Information

human ... CTNNB1(1499), LMNB1(4001)

OMIM accession no.

102582

OMIM accession no.

600555

OMIM accession no.

OMIM accession no.

116806, 150340

storage temp.

−196°C

storage temp.

−196°C

storage temp.

−196°C

storage temp.

−196°C

General description

A549 RFP-STAT3 are lung carcinoma, epithelial cells from a human 58 year old male caucasion having a ZFN modification creating an RFP-STAT3 transgene expressed from the endogenous STAT3 gene locus.

This cell line was derived from ATCC catalog No. CCL-185.

Application

This product is a human A549 cell line in which the genomic STAT3 gene has been endogenously tagged with a Red Fluorescent Protein (RFP) gene using CompoZr® zinc finger nuclease technology. Integration resulted in endogenous expression of the fusion protein in which RFP is attached to the N-terminus of STAT3. Fluorescence imaging shows RFP-STAT3 expression throughout the cell. Upon activation with the ligand IL-6, the cell line shows redistribution of STAT3 to the nucleus, making it useful for high content screening of compounds that modulate STAT3 activity. For example, the redistribution can be abolished by a selective inhibitor of STAT3, Stattic. 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.

To learn more, please visit the Cellular Reporter Cell Line webpage

Features and Benefits

Zinc Finger Nuclease (ZFN)-mediated targeted integration a fluorescent RFP tag just behind the start codon of the STAT3 gene on chromosome 17q21.2 to create a cell line exhibiting stable expression of the transgene tagged with GFP on the N-terminus of the protein.

These A549 cells are adherent with a doubling time of approx. 22 hours.

Quality

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.

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. 1:3-1:6

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% and L-glutamine, Cat. No. G7513, at a final concentration of 2 mM.

Cell freezing medium-DMSO 1X, Cat. No. C6164.

Legal Information

CompoZr is a registered trademark of Merck KGaA, Darmstadt, Germany

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.

Storage Class

10 - Combustible liquids

wgk_germany

WGK 3

flash_point_f

Not applicable

flash_point_c

Not applicable


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