Protocol abstract
Nature Protocols 3, - 606 - 611 (2008)
Published online: 20 March 2008 | doi:10.1038/nprot.2008.18
Subject Categories: Cell and tissue culture | Genetic modification | Imaging | Immunological techniques | Model organisms
Culture of Drosophila S2 cells and their use for RNAi-mediated loss-of-function studies and immunofluorescence microscopy
Stephen L Rogers1,2,3 & Gregory C Rogers1
Abstract
Cultured Drosophila cell lines have become an increasingly popular model system for cell biological and functional genomic studies. One of the most commonly used lines, S2 cells, is particularly useful as it is easy to grow and maintain in the lab, is highly susceptible to gene inhibition using RNAi and is well suited to high-resolution light microscopic assays. Here, we provide protocols for the routine culture and RNAi treatment of S2 cells and methods to prepare these cells for fluorescence microscopy. Using these techniques, loss-of-function experiments may be performed after 4–7 d of RNAi-mediated protein depletion.
- Department of Biology, The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27515, USA.
- Carolina Center for Genome Sciences, The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27515, USA.
- Lineberger Cancer Center, The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27515, USA.
Correspondence to: Stephen L Rogers1,2,3 e-mail: srogers@bio.unc.edu
nature-products
A-Z product listing
- Concanavalin A(MP Biomedicals)
- Coverglass rack, porcelain(Thomas Scientific)
- DEPC-treated water(Invitrogen)
- DMSO(Sigma)
- dNTP mix(Promega)
- Fluorescent mounting medium(Dakocytomation)
MORE ARTICLES LIKE THIS
These links to content published by NPG are automatically generated.
NEWS AND VIEWS
Both a ?magic bullet? and good aim are required to link public health interests and health care needs in HIV infectionNature Medicine News and Views (01 Mar 2000)
