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Methods Mol Biol. 2016;1400:261-80. doi: 10.1007/978-1-4939-3372-3_17.

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

Author information

1
Department of Pathology, Johns Hopkins University School of Medicine, 733 N. Broadway, MRB 469, Baltimore, MD, 21205, USA.
2
Department of Pathology, Johns Hopkins University School of Medicine, 733 N. Broadway, MRB 469, Baltimore, MD, 21205, USA. kburns@jhmi.edu.
3
Department of Pathology, Johns Hopkins University School of Medicine, 733 N. Broadway, MRB 469, Baltimore, MD, 21205, USA. mstaylor@partners.org.
4
Department of Pathology, Massachusetts General Hospital, Boston, MA, USA. mstaylor@partners.org.

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.

KEYWORDS:

Biomarker; Immunofluorescence; Immunohistochemistry; LINE-1 ORF1p; Tissue microarray; Tumor marker; p40

PMID:
26895059
PMCID:
PMC5679191
DOI:
10.1007/978-1-4939-3372-3_17
[Indexed for MEDLINE]
Free PMC Article

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