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CLL1037 Sigma-Aldrich

U2OS GFP-ACTB RFP-TUBA1B

  •  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
  602530
storage temp.   −196°C
Gene Information   human ... ACTB(60), TUBA1B(10376)

Description

General description

This product was derived from ATCC Cat. No. HTB-96.

U2OS GFP-ACTB RFP-TUBA1B are osteosarcoma, epithelial cells, from a human caucasian female (aged 15 years), with GFP-tagged and RFP-tagged modifications.

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 and in which the genomic TUBA1B 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 GFP is attached to the N-terminus of actin and of a fusion protein in which RFP is attached to the N-terminus of tubulin. Fluorescence imaging shows that the actin can polymerize to form the characteristic pattern of actin filaments and that the tubulin can polymerize to form the characteristic pattern of microtubules. 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

TUBA1B (tubulin α 1b) is a microtubule protein, which has an important role in the dynamic process of polymerization and depolymerization during cell replication and division. TUBA1B is overexpressed in hepatocellular carcinoma (HCC) tumor tissues and proliferating 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.

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

ACTB is GFP-tagged on chromosome 7p22.1 and TUBA1B is RFP-tagged on chromosome 12q13.12.

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

Components

U2OS GFP-ACTB RFP-TUBA1B 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

RIDADR 
UN 3245 9
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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