Immunodetection of human LINE-1 expression in cultured cells and human tissues

Reema Sharma, Nemanja Rodić, Kathleen H. Burns, Martin S. Taylor

Research output: Chapter in Book/Report/Conference proceedingChapter

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 specifi c for human LINE-1 ORF1p and its application in immunofl uorescence and immunohistochemistry of both cells and human tissues. We also describe detection of tagged LINE-1 ORF2p via immunofl uorescence. These general methods may be readily adapted to use with many other proteins and antibodies.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages261-280
Number of pages20
DOIs
StatePublished - 2016

Publication series

NameMethods in Molecular Biology
Volume1400
ISSN (Print)1064-3745

Keywords

  • Biomarker
  • Immunofl uorescence
  • Immunohistochemistry
  • LINE-1 ORF1p
  • P40
  • Tissue microarray
  • Tumor marker

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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