U2OS BFP-LMNB1 RFP-ACTB,

Application

This product is a human U2OS cell line in which the genomic LaminB1 gene has been endogenously tagged with a Blue Fluorescent Protein (BFP) and the genomic ACTB gene has been endogenously tagged with a Red Fluorescent Protein (RFP) gene using CompoZr^® Zinc Finger Nuclease technology. Integration resulted in endogenous expression of a fusion protein in which BFP is attached to the N-terminus of LaminB1 and of a fusion protein in which RFP is attached to the N-terminus of actin. Fluorescence imaging shows the characteristic LaminB1 nuclear expression and that the actin can polymerize to form the characteristic pattern of actin filaments. 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.

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Components

U2OS BFP-LMNB1 RFP-ACTB cells

Features and Benefits

LMNB1 is BFP-tagged on chromosome 5q23.3-q31.1 and ACTB is RFP-tagged on chromosome 7p22.1. U2OS cells are adherent, with a doubling time of approx. 29 hours.

General description

This product was derived from ATCC Cat. No. HTB-96.U2OS BFP-LMNB1 RFP-ACTB are osteosarcoma, epithelial cells, from a human caucasian female (aged 15 years), with BFP-tagged and RFP-tagged modifications.

Legal Information

CompoZr is a registered trademark of Sigma-Aldrich Co. LLC

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. 1:3-1:6The base medium for this cell line is McCoy′s 5A1 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 of 10%.The cell freezing medium is DMSO 1X (Cat. No. C6164).

Quality

Tested for Mycoplasma, sterility, post-freeze viability, short terminal repeat (STR) analysis for cell line identification, and PCR assay for cell line species confirmation.