Establishment of a Microplate-Formatted Cell-Based Immunoassay for Rapid Analysis of Nucleotide Excision Repair Ability in Human Primary Cells
Article first published online: 25 JAN 2012
© 2012 Wiley Periodicals, Inc. Photochemistry and Photobiology © 2012 The American Society of Photobiology
Photochemistry and Photobiology
Volume 88, Issue 2, pages 356–362, March/April 2012
How to Cite
Nishinaga, M., Kurata, R., Onishi, K., Kuriyama, K., Wakasugi, M. and Matsunaga, T. (2012), Establishment of a Microplate-Formatted Cell-Based Immunoassay for Rapid Analysis of Nucleotide Excision Repair Ability in Human Primary Cells. Photochemistry and Photobiology, 88: 356–362. doi: 10.1111/j.1751-1097.2012.01073.x
- Issue published online: 1 MAR 2012
- Article first published online: 25 JAN 2012
- Accepted manuscript online: 5 JAN 2012 03:50AM EST
- Received 24 November 2011, accepted 27 December 2011
Figure S1. Human primary keratinocytes were pulse-labeled with 20 mM 5’-bromo-2’deoxyuridine (BrdU) for 30 min. The Cells were fixed with 4% formaldehyde at room temperature for 15 min and subsequently permeabilized with ice-cold 0.5% TritonX-100 in 10 mM PBS (pH 7.4) on ice for 5 min. After denaturing cellular DNA with 2 M HCl at room temperature for 30 min, the cells were stained with anti-BrdU antibody conjugated with Alexa Fluor-594 (Invitrogen), and counterstained with 4’,6-diamidino-2-phenylindole dihydrochloride (DAPI) (Invitrogen). To obtain fluorescence images, BIOREVO BZ-9000 fluorescence microscope (Keyence) was used.
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