Detection of DNA cleavage in apoptotic cells.
Methods Enzymol
; 322: 3-15, 2000.
Article
em En
| MEDLINE
| ID: mdl-10914000
ABSTRACT
At least two discrete deoxyribonuclease activities can be detected during apoptotic death, one that generates 30- to 500-kilobase pair (kbp) domain-sized fragments and another that mediates internucleosomal DNA degradation. The latter nuclease has been identified as the caspase-activated deoxyribonuclease (CAD)/CPAN, a unique enzyme that is normally inhibited by the regulatory subunit ICAD (inhibitor of CAD)/DFF45 (DNA fragmentation factor). In this chapter, techniques widely used to detect DNA cleavage in apoptotic cells, including pulsed-field gel electrophoresis, conventional agarose gel electrophoresis, and terminal transferase-mediated dUTP nick end-labeling (TUNEL), are briefly reviewed. In addition, the use of ICAD to inhibit apoptosis-associated nuclease activity is illustrated. When properly applied, these techniques are widely applicable to the characterization of apoptotic cells.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
DNA
/
Apoptose
Tipo de estudo:
Diagnostic_studies
/
Prognostic_studies
Limite:
Humans
Idioma:
En
Revista:
Methods Enzymol
Ano de publicação:
2000
Tipo de documento:
Article
País de afiliação:
Estados Unidos