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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.

Original languageEnglish
Title of host publicationMethods in Enzymology
EditorsNancy L. Allbritton, Michelle L. Kovarik
Number of pages15
PublisherElsevier
Publication year1 Jan 2019
Pages43-57
ISBN (print)9780128170908
DOIs
Publication statusPublished - 1 Jan 2019
SeriesMethods in Enzymology
Volume628
ISSN0076-6879

    Research areas

  • Cancer, Cancer stem cells, Enzyme activity, Human topoisomerase 1, Microscopic slides, Rolling circle amplification, Single cell detection

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