U2OS GFP-TUBA1B,

Application

To learn more, please visit the Cellular Reporter Cell Line webpage

This product is a human U2OS cell line in which the genomic TUBA1B 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 tubulin. Fluorescence imaging shows that the tubulin can polymerize to form the characteristic pattern of microtubules. Addition of vincristine to this cell line shows the expected depolymerization 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.

Biochem/physiol Actions

K-α1 tubulin/ tubulin α 1b (TUBA1B) participates in several cellular activities like GTP binding, GTPase activity, maintaining cellular structure in the form of microtubules and microtubule-based intracellular movement. It plays a crucial role in polymerization and depolymerization during cell replication and division. TUBA1B is overexpressed in hepatocellular carcinoma (HCC) tumor tissues and proliferating HCC cells.

Tubulin α 1b (TUBA1B) plays an important role in dynamic process of polymerization and depolymerization during cell replication and division. Elevated expression of the gene has been observed 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.

Components

U2OS GFP-TUBA1B cells

Features and Benefits

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

GFP-tagged on chromosome 12q13.12

General description

K-α1 tubulin/ tubulin α 1b (TUBA1B) is a microtubule protein. It is a ~50 kDa protein in its poly glycosylated, post-translational form. TUBA1B is present in cells as one of the six different isotypes. This gene is located on human chromosome 12q13.12.

This product corresponds to ATCC Cat. No. HTB-96.

U2OS GFP-TUBA1B 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 Sigma-Aldrich Co. LLC

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. 1:3-1:6The 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).

Quality

Tested for Mycoplasma, sterility, post-freeze viability, short terminal repeat (STR) analysis for cell line identification, PCR assay for cell line species confirmation.