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CLL1218

Sigma-Aldrich

U2OS LMNB1-TUBA1B-ACTB

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NACRES:
NA.81

biological source

human female bone (Source Disease: Osteosarcoma)

OMIM accession no.

storage temp.

−196°C

Gene Information

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U2OS RFP-TUBA1B

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CLL1034

U2OS RFP-TUBA1B

U2OS Cells GFP-NUP98

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CLL1136

U2OS Cells GFP-NUP98

U2OS RFP-ACTB

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CLL1035

U2OS RFP-ACTB

OMIM accession no.

102630, 150340, 602530

OMIM accession no.

602530

OMIM accession no.

601021

OMIM accession no.

102630

storage temp.

−196°C

storage temp.

−196°C

storage temp.

−196°C

storage temp.

−196°C

Quality Level

200

Quality Level

200

Quality Level

200

Quality Level

200

Gene Information

human ... ACTB(60), LMNB1(4001), TUBA1B(10376)

Gene Information

human ... alpha tubulin 1B(10376)

Gene Information

human ... NUP98(4928)

Gene Information

human ... beta actin(60)

General description

U2OS LMNB1-TUBA1B-ACTB are bone osteosarcoma cells from a human 15 year old caucasion female having three distinct ZFN modifications creating a BFP-tagged LMNB1 transgene, a GFP-tagged TUBA1B transgene and a RFP-tagged ACTB transgene expressed from their endogenous gene locus.

This cell line was derived from ATCC Catalog No. HTB-96.

Application

This product is a human U2OS cell line in which three genomic loci - LMNB1, TUBA1B, and ACTB - have been endogenously tagged with fluorescent protein genes for Blue Fluorescent Protein (BFP), Green Fluorescent Protein (GFP), and Red Fluorescent Protein (RFP), respectively, using CompoZr® zinc finger nuclease (ZFN) technology. Integration resulted in endogenous expression of: 1) the fluorescent fusion protein, BFP-laminB1, easily detected in the nuclear envelope; 2) the fluorescent fusion protein, GFP-α-tubulin, that polymerizes to form characteristic patterns of microtubules; 3) the fluorescent fusion protein, RFP-β-actin, that polymerizes to form characteristic patterns of actin fibers. Imaging of the cell line shows fluorescent labeling of the three proteins having normal cellular distribution patterns which makes the cell line ideal for high content screening to identify compounds that modulate cellular activity. This stable cell line was expanded from a single cell clone. The LMNB1, TUBA1B, and ACTB gene regulation and corresponding protein function are preserved in contrast to cell lines exhibiting overexpression from an exogenous promoter.
To learn more, please visit the Cellular Reporter Cell Line webpage

Biochem/physiol Actions

Nuclear lamin B1, encoded by the LMNB1 gene, is an intermediate filament protein of the nuclear envelope. The protein exhibits transcriptional co-regulatory activity and plays a key role in DNA replication, cellular aging and stress responses. Thus, LMNB1 can be used as a potential biomarker for HCC.
Tubulin α 1b (TUBA1B) is a microtubule protein, which plays a vital role in dynamic process of polymerization and depolymerization during cell replication and division. TUBA1B is overexpressed in hepatocellular carcinoma (HCC) tumor tissues and proliferating HCC cells. This upregulated expression of the gene leads to poor prognosis and chemotherapy resistance in HCC.
Actin β (ACTB) is an abundant cytoskeletal housekeeping protein. The protein is expressed in the nucleus and controls gene expression, cell division and proliferation. ACTB is generally used as a reference gene in measuring expression levels in tumors.

Features and Benefits

Zinc Finger Nuclease (ZFN)-mediated targeted integration of three different fluorescent tags: LMNB1 is BFP-tagged on chromosome 5q23.2-q31.1, TUBA1B is GFP-tagged on chromosome 12q13.12 and ACTB is RFP-tagged on chromosome 7p22.1 to create a cell line exhibiting stable expression of all three transgenes.

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

Quality

Tested for Mycoplasma, sterility, post-freeze viability, short terminal repeat (STR) analysis for cell line identification, cytochrome oxidase I (COI) analysis 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 5A 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 (v/v) of 10%.

Cell freezing medium-DMSO 1X, Cat. No. C6164.

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.

Storage Class Code

10 - Combustible liquids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


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Customers Also Viewed

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Structural Organization of the Human Gene (LMNB1) Encoding Nuclear Lamin B1
Lin F, et al.
Genomics, 27(2), 230-236 (1995)
ACTB in cancer.
Guo C, et al.
Clinica Chimica Acta; International Journal of Clinical Chemistry, 417, 39-44 (2013)
Barney Boyce et al.
Scientific reports, 8(1), 14755-14755 (2018-10-05)
As an alternative to laser-based methods, we developed a novel in situ cell isolation method and instrument based on local water absorption of millimeter wave (MMW) radiation that occurs in cellular material and nearby culture medium while the cultureware materials
Circulating Lamin B1 (LMNB1) Biomarker Detects Early Stages of Liver Cancer in Patients
Sun S, et al.
Journal of Proteome Research, 9(1), 70-78 (2009)
Increased α-tubulin1b expression indicates poor prognosis and resistance to chemotherapy in hepatocellular carcinoma.
Lu C, et al.
Digestive Diseases and Sciences, 58(9), 2713-2720 (2013)

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