MCF10A RFP-TUBA1B,

Application

MCF10A RFP-TUBA1B has been used to study the role of BRCA1 on the cell division axis.

MCF10A RFP-TUBA1B has been used in cell culture assays and western blot analysis.

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

This product is a human MCF10A cell line 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 the fusion protein in which RFP 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 paclitaxel causes the expected bundling 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) 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.

Components

MCF10A RFP-TUBA1B cells

Features and Benefits

RFP-tagged on chromosome 12q13.12.MCF10A cells are adherent, with a doubling time of approx.20 hours.

General description

This product was derived from ATCC Cat. No. CRL-10317.MCF10A RFP-TUBA1B are fibrocystic disease, epithelial cells, from a human female (aged 36 years), with a RFP-tagged modification.

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.

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 the vial by gentle agitation in a 37°C water bath (~2 minutes), keeping the 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 to 1:4The base medium for this cell line is Dulbecco′s Modified Eagle′s Medium/Ham′s Nutrient Mixture F-12, Cat. No. 51448C. To make the complete growth medium, add the following components to the base medium: horse serum, Cat. No. H1270, to a final concentration (v/v) of 5%; cholera toxin, Cat. No. C8052, to a final concentration of 1 ng/mL, human insulin, Cat. No. I9278, to a final concentration of 10 µg/mL, epidermal growth factor, Cat. No. E9644, to a final concentration of 10 ng/mL; and hydrocortisone, Cat. No. H6909, to a final concentration of 0.5 µg/mL.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.