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Detection of Isoforms Differing by a Single Charge Unit in Individual Cells
Author(s) -
Tentori Augusto M.,
Yamauchi Kevin A.,
Herr Amy E.
Publication year - 2016
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201606039
Subject(s) - gene isoform , isoelectric focusing , cytoplasm , lysis , chemistry , isoelectric point , resolution (logic) , cell , single cell analysis , biophysics , selectivity , biochemistry , microbiology and biotechnology , biology , enzyme , gene , computer science , artificial intelligence , catalysis
To measure protein isoforms in individual mammalian cells, we report single‐cell resolution isoelectric focusing (scIEF) and high‐selectivity immunoprobing. Microfluidic design and photoactivatable materials establish the tunable pH gradients required by IEF and precisely control the transport and handling of each 17‐pL cell lysate during analysis. The scIEF assay resolves protein isoforms with resolution down to a single‐charge unit, including both endogenous cytoplasmic and nuclear proteins from individual mammalian cells.