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High-throughput subcellular protein localization using transfected-cell arrays : subcellular protein localization using cell arrays

Item Type:Article
Title:High-throughput subcellular protein localization using transfected-cell arrays : subcellular protein localization using cell arrays
Creators Name:Hu, Y. and Janitz, M.
Abstract:Knowledge of protein localization within the cellular environment is critical for understanding the function of the protein and its regulatory networks. Protein localization data, however, have traditionally been accumulated from single or small-scale experiments. Transfected-cell arrays (TCAs) represent a robust alternative for the high-throughput analysis of gene/protein functions in mammalian cells. For protein localization studies, TCAs not only allow for the transfection and expression of over 1,000 genes in a single experiment but also make it feasible for simultaneous co-localization analyses of different subcellular compartments. In this chapter, we have described a protein co-localization protocol using transfected human cell arrays for a large set of cellular compartments, including the nucleus, ER, Golgi apparatus, mitochondrion, lysosome, peroxisome, and the microtubules, intermediate filaments and actin filaments. The application of these "organelle-co-localized cell arrays" facilitates the precise determination of the localizations of numerous recombinant proteins in a single experiment, making it currently the most efficient technique for high-throughput protein co-localization screening.
Keywords:Transfected-Cell Array, Protein Localization, Organelle Counterstaining, Reverse Transfection, Confocal Immunofluorescence, His6-Tag
Source:Methods in Molecular Biology
ISSN:1064-3745
Publisher:Springer / Humana Press (U.S.A.)
Volume:706
Page Range:53-72
Date:2011
Official Publication:https://doi.org/10.1007/978-1-61737-970-3_5
PubMed:View item in PubMed

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