Format

Send to:

Choose Destination
See comment in PubMed Commons below
Methods Enzymol. 2013;529:209-26. doi: 10.1016/B978-0-12-418687-3.00017-3.

Generating mammalian stable cell lines by electroporation.

Author information

  • 1Johns Hopkins University School of Medicine, Baltimore, MD, USA.

Abstract

Expression of functional, recombinant mammalian proteins often requires expression in mammalian cells (see Single Cell Cloning of a Stable Mammalian Cell Line). If the expressed protein needs to be made frequently, it can be best to generate a stable cell line instead of performing repeated transient transfections into mammalian cells. Here, we describe a method to generate stable cell lines via electroporation followed by selection steps. This protocol will be limited to the CHO dhfr-Urlaub et al. (1983) and LEC1 cell lines, which in our experience perform the best with this method.

Copyright © 2013 Elsevier Inc. All rights reserved.

KEYWORDS:

Colonies of cells; Dilute plasmid DNA; Electroporation; Methotrexate (MTX); Plating electroporated cells; Protocol

PMID:
24011048
[PubMed - indexed for MEDLINE]
PMCID:
PMC4088271
Free PMC Article
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for Elsevier Science Icon for PubMed Central
    Loading ...
    Write to the Help Desk