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CLL1032 Sigma-Aldrich

U2OS GFP-ACTB

  •  NACRES NA.81

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Properties

Related Categories Cell Biology, Cellular Reporters, CompoZr Cell Lines More...
Quality Level   200
biological source   human female bone (Source Disease: Osteosarcoma)
OMIM accession no.   102630
storage temp.   −196°C
Gene Information   human ... beta actin(60)

Description

General description

This product was derived from ATCC Cat. No. HTB-96.
ACTB (actin beta) gene codes for β-actin. It is a cytoskeletal housekeeping protein. It is present in the cytoplasm. The ACTB gene is located on human chromosome 7p22.1. GFP (green fluorescent protein) is a major reporter protein that has fluorescence properties. It can fluoresce without the help of any cofactors or substrates except for oxygen.

U2OS GFP-ACTB are osteosarcoma, epithelial cells, from a human caucasian female (aged 15 years), with a GFP-tagged modification.

Application

This product is a human U2OS cell line in which the genomic ACTB gene has been endogenously tagged with a Green Fluorescent Protein (GFP) gene using CompoZr® Zinc Finger Nuclease technology. Integration resulted in endogenous expression of the fusion protein in which GFP is attached to the N-terminus of actin. Fluorescence imaging shows 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.

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

Biochem/physiol Actions

Actin β (ACTB) is an abundant cytoskeletal housekeeping protein. The protein, expressed in the nucleus, controls gene expression, cell division and proliferation. ACTB is generally used as a reference gene in measuring expression levels in tumors.

β-Actin modulates cell growth, migration and the G-actin pool. Mutations in ACTB leads to pleiotropic developmental disorder. GFP (green fluorescent protein) is used for quantitative analysis.

Features and Benefits

GFP-tagged on chromosome 7p22.1

The U2OS cells are adherent, with a doubling time of approx. 29 hours.

Components

U2OS GFP-ACTB cells

Quality

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

Legal Information

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

Safety & Documentation

Safety Information

WGK Germany 
WGK 3
Flash Point(F) 
Not applicable
Flash Point(C) 
Not applicable
Protocols & Articles
Peer-Reviewed Papers
15

References

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