Immunodetection of Human LINE-1 Expression in Cultured Cells and Human Tissues.
Methods Mol Biol
; 1400: 261-80, 2016.
Article
em En
| MEDLINE
| ID: mdl-26895059
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.
Palavras-chave
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Imuno-Histoquímica
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Expressão Gênica
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Fases de Leitura Aberta
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Imunofluorescência
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Elementos Nucleotídeos Longos e Dispersos
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Análise Serial de Tecidos
Limite:
Humans
Idioma:
En
Revista:
Methods Mol Biol
Assunto da revista:
BIOLOGIA MOLECULAR
Ano de publicação:
2016
Tipo de documento:
Article
País de afiliação:
Estados Unidos