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Improved method to retain cytosolic reporter protein fluorescence while staining for nuclear proteins
Author(s) -
Heinen André P.,
Wanke Florian,
Moos Sonja,
Attig Sebastian,
Luche Hervé,
Pal Prajna Paramita,
Budisa Nediljko,
Fehling Hans Jörg,
Waisman Ari,
Kurschus Florian C.
Publication year - 2014
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.22451
Subject(s) - staining , cytosol , fluorescence , microbiology and biotechnology , flow cytometry , green fluorescent protein , fluorescent staining , protein subcellular localization prediction , biology , chemistry , biophysics , biochemistry , gene , genetics , physics , quantum mechanics , enzyme
Staining of transcription factors (TFs) together with retention of fluorescent reporter proteins is hindered by loss of fluorescence using current available methods. In this study, it is shown that current TF staining protocols do not destroy fluorescent proteins (FPs) but rather that fixation is not sufficient to retain FPs in the cytosol of the permeabilized cells. In this article, a simple and reliable protocol is elaborated, which allows efficient TF and cytokine staining while retaining FPs inside fixed cells. © 2014 International Society for Advancement of Cytometry