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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
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.
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.
Features and Benefits
GFP-tagged on chromosome 7p22.1
The U2OS cells are adherent, with a doubling time of approx. 29 hours.
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).
Analysis Note
Tested for Mycoplasma, sterility, post-freeze viability, short terminal repeat (STR) analysis for cell line identification, PCR assay for cell line species confirmation.
Other Notes
U2OS GFP-ACTB cells
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.
| biological source | human female bone (Source Disease: Osteosarcoma) |
| Quality Segment | 200 |
| OMIM accession no. | 102630 |
| storage temp. | −196°C |
| Gene Information | human ... beta actin(60) |
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