U2OS GFP-ACTB
SIGMA/CLL1032
Product Type: Chemical
Fluorescence Microscopy U2OS GFP-ACTB: Cat. No. CLL1032: U2OS osteosarcoma knock-in cell line expressing the ACTB gene (β-actin) endogenously tagged with GFP at the N-terminus. Cells were imaged live in Hanks Balanced Salt solution, Cat. No. H8264, supplemented with 2% Fetal Bovine Serum, Cat. No. F2442, using GFP filterset (ex 450–490/em 500–550) and 40×/1.3 oil objective.
Differential Interference Contrast (DIC) Microscopy U2OS GFP-ACTB: Cat. No. CLL1032: Differential interference contrast (DIC) microscopy of the same cells seen in the fluorescence microscopy image.
| biological source | human female bone (Source Disease: Osteosarcoma) |
| OMIM accession no. | 102630   |
| Quality Level | 200 |
| storage temp. | −196°C |
| 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. |
| 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. |
| Application: | 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. |
| 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. |
| 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. |
| Features and Benefits: | GFP-tagged on chromosome 7p22.1 |
| Features and Benefits: | The U2OS cells are adherent, with a doubling time of approx. 29 hours. |
| 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. |
| General description: | U2OS GFP-ACTB are osteosarcoma, epithelial cells, from a human caucasian female (aged 15 years), with a GFP-tagged modification. |
| Legal Information: | CompoZr is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Other Notes: | U2OS GFP-ACTB cells |
| 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). |
| WGK Germany | WGK 3 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | −196°C |
| UNSPSC | 41106514 |