Immunonucleochemistry: a new method for in situ detection of antigens in the nucleus of cells in culture.

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

Cytotechnology

journal_title

Cytotechnology

authors

Moon IS,Lee H,Park SD,Seog DH

doi

10.1007/s10616-010-9266-0

subject

Has Abstract

pub_date

2010-04-01 00:00:00

pages

83-93

issue

2

eissn

0920-9069

issn

1573-0778

journal_volume

62

pub_type

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