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Nat Methods. 2017 Aug;14(8):789-792. doi: 10.1038/nmeth.4342. Epub 2017 Jun 26.

A DNA origami platform for quantifying protein copy number in super-resolution.

Author information

1
ICFO-Institut de Ciencies Fotoniques, The Barcelona Institute of Science and Technology, Castelldefels, Spain.
2
Universitat de Vic - Universitat Central de Catalunya (UVic-UCC), Vic, Spain.
3
Department of Biological Sciences, Smith College, Northampton, Massachusetts, USA.
4
Center for Microscopy and Imaging, Smith College, Northampton, Massachusetts, USA.
5
ICREA, Barcelona, Spain.

Abstract

Single-molecule-based super-resolution microscopy offers researchers a unique opportunity to quantify protein copy number with nanoscale resolution. However, while fluorescent proteins have been characterized for quantitative imaging using calibration standards, similar calibration tools for immunofluorescence with small organic fluorophores are lacking. Here we show that DNA origami, in combination with GFP antibodies, is a versatile platform for calibrating fluorophore and antibody labeling efficiency to quantify protein copy number in cellular contexts using super-resolution microscopy.

PMID:
28650478
PMCID:
PMC5534338
DOI:
10.1038/nmeth.4342
[Indexed for MEDLINE]
Free PMC Article

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