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  • A chemical method for fast and sensitive detection of DNA synthesis in vivo.

A chemical method for fast and sensitive detection of DNA synthesis in vivo.

Proceedings of the National Academy of Sciences of the United States of America (2008-02-15)
Adrian Salic, Timothy J Mitchison
ABSTRACT

We have developed a method to detect DNA synthesis in proliferating cells, based on the incorporation of 5-ethynyl-2'-deoxyuridine (EdU) and its subsequent detection by a fluorescent azide through a Cu(I)-catalyzed [3 + 2] cycloaddition reaction ("click" chemistry). Detection of the EdU label is highly sensitive and can be accomplished in minutes. The small size of the fluorescent azides used for detection results in a high degree of specimen penetration, allowing the staining of whole-mount preparations of large tissue and organ explants. In contrast to BrdU, the method does not require sample fixation or DNA denaturation and permits good structural preservation. We demonstrate the use of the method in cultured cells and in the intestine and brain of whole animals.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
5-Ethynyl-2′-deoxyuridine, 95%
Sigma-Aldrich
In Vivo EdU Flow Cytometry 50 Kit 488, sufficient for 50 assays
Sigma-Aldrich
In Vivo EdU Flow Cytometry Kit 647, sufficient for 50 assays
Sigma-Aldrich
In Vivo EdU Flow Cytometry 50 Kit 488, sufficient for 50 assays
Sigma-Aldrich
In Vivo EdU Flow Cytometry Kit 594, sufficient for 50 assays
Sigma-Aldrich
In Vivo EdU Flow Cytometry Kit 647, sufficient for 50 assays
Sigma-Aldrich
In Vivo EdU Flow Cytometry Kit 594, sufficient for 50 assays
Sigma-Aldrich
In Vivo EdU Flow Cytometry Kit 647, sufficient for 50 assays
Sigma-Aldrich
In Vivo EdU Flow Cytometry Kit 594, sufficient for 50 assays
Sigma-Aldrich
In Vivo EdU HTS 2x96 Kit 555, sufficient for 200 assays
Sigma-Aldrich
In Vivo EdU HTS 2x96 Kit 488, sufficient for 200 assays
Sigma-Aldrich
In Vivo EdU Flow Cytometry Kit 555, sufficient for 50 assays
Sigma-Aldrich
In Vivo EdU HTS 2x96 Kit 555, sufficient for 200 assays
Sigma-Aldrich
In Vivo EdU Flow Cytometry Kit 555, sufficient for 50 assays
Sigma-Aldrich
In Vivo EdU HTS 2x96 Kit 488, sufficient for 200 assays
Sigma-Aldrich
In Vivo EdU Flow Cytometry Kit 555, sufficient for 50 assays