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Immunodetection of Human LINE-1 Expression in Cultured Cells and Human Tissues

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Transposons and Retrotransposons

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1400))

Abstract

Long interspersed element-1 (LINE-1) is the only active protein-coding retrotransposon in humans. It is not expressed in somatic tissue but is aberrantly expressed in a wide variety of human cancers. ORF1p protein is the most robust indicator of LINE-1 expression; the protein accumulates in large quantities in cellular cytoplasm. Recently, monoclonal antibodies have allowed more complete characterizations of ORF1p expression and indicated potential for developing ORF1p as a clinical biomarker. Here, we describe a mouse monoclonal antibody specific for human LINE-1 ORF1p and its application in immunofluorescence and immunohistochemistry of both cells and human tissues. We also describe detection of tagged LINE-1 ORF2p via immunofluorescence. These general methods may be readily adapted to use with many other proteins and antibodies.

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Acknowledgement

We thank Carolyn Machamer and Travis Ruch for their immunofluorescence expertise. We thank Jeff Han for the gift of rabbit anti-ORF1 JH73. We thank Norman Barker for photography of IHC slides. Funding for these projects has been provided in part by R01CA163705 (KHB) and the Burroughs Wellcome Fund Career Awards for Medical Scientists (KHB).

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Authors and Affiliations

  1. Department of Pathology, Johns Hopkins University School of Medicine, 733 N. Broadway, MRB 469, Baltimore, MD, 21205, USA

    Reema Sharma, Nemanja Rodić, Kathleen H. Burns M.D., Ph.D. & Martin S. Taylor

  2. Department of Pathology, Massachusetts General Hospital, Boston, MA, USA

    Martin S. Taylor

Authors
  1. Reema Sharma
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  2. Nemanja Rodić
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  3. Kathleen H. Burns M.D., Ph.D.
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  4. Martin S. Taylor
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Corresponding author

Correspondence to Kathleen H. Burns M.D., Ph.D. .

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Editors and Affiliations

  1. Institute of Genetics and Molecular Medi, Genyo (Center for Genomics and Oncologic, Granada, Spain

    Jose L. Garcia-Pérez

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Sharma, R., Rodić, N., Burns, K.H., Taylor, M.S. (2016). Immunodetection of Human LINE-1 Expression in Cultured Cells and Human Tissues. In: Garcia-Pérez, J. (eds) Transposons and Retrotransposons. Methods in Molecular Biology, vol 1400. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3372-3_17

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  • DOI: https://doi.org/10.1007/978-1-4939-3372-3_17

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3370-9

  • Online ISBN: 978-1-4939-3372-3

  • eBook Packages: Springer Protocols

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