Abstract:
:The advancement of immunocytochemistry (ICC) allows one to observe detailed spatial distribution of cellular antigens, but, with some limitations. Using conventional ICC, it is difficult to distinguish the nuclear localization from cytoplasm, as two large subcellular compartments overlap on the z-axis. In this study, we have investigated whether in situ immunostaining of 'naked' nuclei could provide an unambiguous method for detection of nuclear antigens. We have designed a protocol that efficiently lyses plasmalemma, while keeping the nuclear envelope intact. The optimal condition for lysing the plasmalemma was 0.5% Nonidet P-40 for 5 min in both neuronal and non-neuronal cultured cells. Using this protocol, we could unambiguously isolate nuclear from cytoplasmic ICC signals. Since the present protocol has been designed for immunostaining of 'naked' nuclei from cultured or isolated cells, we have coined a new term to refer to this procedure as 'immunonucleochemistry' ('INC' for abbreviation).
journal_name
Cytotechnologyjournal_title
Cytotechnologyauthors
Moon IS,Lee H,Park SD,Seog DHdoi
10.1007/s10616-010-9266-0subject
Has Abstractpub_date
2010-04-01 00:00:00pages
83-93issue
2eissn
0920-9069issn
1573-0778journal_volume
62pub_type
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