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Enzymatic activity in single cells

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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.

TitelMethods in Enzymology
RedaktørerNancy L. Allbritton, Michelle L. Kovarik
Antal sider15
Udgivelsesår1 jan. 2019
ISBN (trykt)9780128170908
StatusUdgivet - 1 jan. 2019
SerietitelMethods in Enzymology

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