Chapter title |
Immunodetection of Human LINE-1 Expression in Cultured Cells and Human Tissues.
|
---|---|
Chapter number | 17 |
Book title |
Transposons and Retrotransposons
|
Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-3372-3_17 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3370-9, 978-1-4939-3372-3
|
Authors |
Reema Sharma, Nemanja Rodić, Kathleen H. Burns M.D., Ph.D., Martin S. Taylor, Kathleen H. Burns |
Editors |
Jose L. Garcia-Pérez |
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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