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CLL1136

Sigma-Aldrich

U2OS Cells GFP-NUP98

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NACRES:
NA.81

biological source

human female bone (Source Disease: Bone osteosarcoma)

Quality Level

OMIM accession no.

storage temp.

−196°C

Gene Information

human ... NUP98(4928)

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U2OS Cells GFP-NUP98

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CLL1036

U2OS GFP-HMGA1

vibrant-m

CLL1038

U2OS BFP-LMNB1 RFP-ACTB

vibrant-m

CLL1034

U2OS RFP-TUBA1B

storage temp.

−196°C

storage temp.

−196°C

storage temp.

−196°C

storage temp.

−196°C

Gene Information

human ... NUP98(4928)

Gene Information

human ... HMGA1(3159)

Gene Information

human ... ACTB(60), LMNB1(4001)

Gene Information

human ... alpha tubulin 1B(10376)

Quality Level

200

Quality Level

200

Quality Level

200

Quality Level

200

OMIM accession no.

601021

OMIM accession no.

600701

OMIM accession no.

102630, 150340

OMIM accession no.

602530

General description

U2OS GFP-NUP98 are bone osteosarcoma cells from a human 15 year old caucasian female having a ZFN modification creating a GFP-NUP98 transgene expressed from the endogenous NUP98 gene locus.

This cell line was derived from ATCC Catalog No. HTB-96.

Application

U2OS Cells have been used to assess cell proliferation using Alamar Blue and live cell imaging. This product is a human U2OS cell line in which the genomic NUP98 gene has been endogenously tagged with a 3XFLAG epitope and a Green Fluorescent Protein (GFP) gene using CompoZr® Zinc Finger Nuclease technology. Integration resulted in endogenous expression of a fusion protein in which the 3XFLAG is attached to the N-terminus of GFP and the GFP is attached to the N-terminus of NUP98. Fluorescence imaging shows characteristic NUP98 nuclear expression. 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 of a fluorescent GFP tag just behind the start codon of the NUP98 gene on chromosome 11p15.4 to create a cell line exhibiting stable expression of the transgene tagged with GFP on the N-terminus of the protein.

The U2OS cells are adherent, with a doubling time of approx. 29 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 McCoy′s 5A Medium Modified, Cat. No. M9309. 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%.

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

Legal Information

3xFLAG is a trademark of Sigma-Aldrich Co. LLC
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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PP 60-65 Cell Proliferation Study of Human Osteosarcoma Cell Line (U2OS) using Alamar Blue Assayand Live Cell Imaging
Marahaini Musa
IOSR Journal of Dental and Medical Sciences, 8(2) (2013)

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