title: Immunodetection of Human LINE-1 Expression in Cultured Cells and Human Tissues.
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.
authors.affiliation: Department of Pathology, Johns Hopkins University School of Medicine, 733 N. Broadway, MRB 469, Baltimore, MD, 21205, USA.
authors.forename: Reema
authors.initials: R
authors.lastname: Sharma
authors.affiliation: Department of Pathology, Johns Hopkins University School of Medicine, 733 N. Broadway, MRB 469, Baltimore, MD, 21205, USA.
authors.forename: Nemanja
authors.initials: N
authors.lastname: Rodić
authors.affiliation: Department of Pathology, Johns Hopkins University School of Medicine, 733 N. Broadway, MRB 469, Baltimore, MD, 21205, USA. kburns@jhmi.edu.
authors.forename: Kathleen H
authors.initials: KH
authors.lastname: Burns
authors.affiliation: Department of Pathology, Johns Hopkins University School of Medicine, 733 N. Broadway, MRB 469, Baltimore, MD, 21205, USA. mstaylor@partners.org.|Department of Pathology, Massachusetts General Hospital, Boston, MA, USA. mstaylor@partners.org.
authors.forename: Martin S
authors.initials: MS
authors.lastname: Taylor
mesh_terms: D015415:Biomarkers; D002460:Cell Line; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D020084:Long Interspersed Nucleotide Elements; D016366:Open Reading Frames; D046888:Tissue Array Analysis / Q000379:methods
chemical_list: D015415:Biomarkers
keywords: Biomarker; Immunofluorescence; Immunohistochemistry; LINE-1 ORF1p; Tissue microarray; Tumor marker; p40