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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.

SUBMITTER: Sharma R

PROVIDER: S-EPMC5679191 | biostudies-literature | 2016

REPOSITORIES: biostudies-literature

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

Sharma Reema R Rodić Nemanja N Burns Kathleen H KH Taylor Martin S MS

Methods in molecular biology (Clifton, N.J.) 20160101

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. H ...[more]

PMID: 26895059

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