Enzymatic activity in single cells.
Methods Enzymol
; 628: 43-57, 2019.
Article
em En
| MEDLINE
| ID: mdl-31668235
ABSTRACT
With the increasing recognition of the importance in addressing cell-to-cell variations for the understanding of complex biological systems, single cell analyses are becoming increasingly important. Presented in this chapter is a highly sensitive approach capable of measuring human topoisomerase 1 (TOP1) activity in single CD133 positive DLD-1 cells. The method termed On-Slide "Rolling circle Enhanced Enzyme Activity Detection (REEAD)" relies on the specific capture and lysis of CD133 positive cells on glass slides dual functionalized with anti-CD133 antibodies and a specific DNA primer. The On-Slide REEAD was demonstrated to be directly quantitative. Furthermore, the method allowed for the highly sensitive detection of TOP1 activity in single CD133 positive DLD-1 cells. The described protocol is expected to open for new possibilities in the single cell research, particularly for the investigations of chemoresistance of the cancer stem cells.
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Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
DNA Topoisomerases Tipo I
/
Ensaios Enzimáticos
/
Análise de Célula Única
Limite:
Humans
Idioma:
En
Ano de publicação:
2019
Tipo de documento:
Article