U2OS BFP-LMNB1 RFP-ACTB
SIGMA/CLL1038
Product Type: Chemical
Fluorescence microscopy (overlay) U2OS BFP-LMNB1 RFP-ACTB: Cat. No. CLL1038: U2OS osteosarcoma double knock-in cell line expressing the LMNB1 gene (lamin B1) endogenously tagged with BFP at the N-terminus and the ACTB gene (β-actin) endogenously tagged with RFP 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 BFP filterset (ex 395-410/em 430-480), RFP filterset (ex 530–560/em 590–650) and 40×/1.3 oil objective.
Differential Interference Contrast (DIC) Microscopy U2OS BFP-LMNB1 RFP-ACTB: Cat. No. CLL1038: Differential interference contrast (DIC) microscopy of the same cells seen in the fluorescence microscopy image.
Fluorescence Microscopy U2OS BFP-LMNB1 RFP-ACTB: Cat. No. CLL1038: BFP channel used in the overlay image.
Fluorescence Microscopy U2OS BFP-LMNB1 RFP-ACTB: Cat. No. CLL1038: RFP channel used in the overlay image.
| biological source | human female bone (Disease source: Osteosarcoma) |
| OMIM accession no. | 102630   |
| 150340 |
|
| 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, and PCR assay for cell line species confirmation. |
| 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. |
| Application: | To learn more, please visit the Cellular Reporter Cell Line webpage |
| 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: | 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 Merck KGaA, Darmstadt, Germany |
| Other Notes: | U2OS BFP-LMNB1 RFP-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). |
| RIDADR | UN 3245 9 |
| WGK Germany | WGK 3 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | −196°C |
| UNSPSC | 41106514 |