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

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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).

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Abbreviations

NAGK:

N-acetylglucosamine kinase

DIV:

Day in vitro

HEK:

Human embryonic kidney

ICC:

Immunocytochemistry

IHC:

Immunohistochemistry

INC:

Immunonucleochemistry

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Acknowledgments

We thank Eun-jung Jung for her excellent technical assistance and Dr. Samikshan Dutta for critical reading. This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2009-0076950).

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Authors and Affiliations

  1. Department of Anatomy, College of Medicine, Dongguk University, Gyeongju, 780-714, Korea

    Il Soo Moon & HyunSook Lee

  2. Department of General Thoracic and Cardiovascular Surgery, College of Medicine, Dongguk University, Gyeongju, 780-714, Korea

    Sung Dong Park

  3. Department of Biochemistry, College of Medicine, Inje University, Pusan, 614-735, Korea

    Dae-Hyun Seog

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  1. Il Soo Moon
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  2. HyunSook Lee
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  3. Sung Dong Park
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  4. Dae-Hyun Seog
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Correspondence to Il Soo Moon.

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Moon, I.S., Lee, H., Park, S.D. et al. Immunonucleochemistry: a new method for in situ detection of antigens in the nucleus of cells in culture. Cytotechnology 62, 83–93 (2010). https://doi.org/10.1007/s10616-010-9266-0

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  • DOI: https://doi.org/10.1007/s10616-010-9266-0

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