Single-cell protein analysis of a single mouse embryo by two-dimensional capillary electrophoresis
Single-cell protein analysis of a single mouse embryo by two-dimensional capillary electrophoresis
Available online 19 June 2006.
Melissa M. Harwooda, Elisabeth S. Christiansb, Md. Abul Fazala and Norman J. Dovichia
Journal of Chromatography A
Volume 1130, Issue 2 , 20 October 2006
ScienceDirect
Copyright ? 2006 Elsevier B.V. All rights reserved.
aDepartment of Chemistry, University of Washington, Seattle, WA 98195, USA
bCentre de Biologie du Developpement ? UMR5547, CNRS/Universite Paul Sabatier, 118 Route de Narbonne-bat 4R3B3, 31062 Toulouse, France
Abstract
The characterization of protein expression from a single-cell mouse embryo using two-dimensional capillary electrophoresis (2D-CE) is described. These zygotes were obtained from Hsf1 gene knockout mice. Single zygotes were lysed off-column and proteins were fluorescently labeled using the fluorogenic dye 3-(2-furoyl)quinoline-2-carboxaldehyde (FQ). After injection, analytes were separated first according to molecular weight using capillary sieving electrophoresis (CSE) and then by micellar electrokinetic capillary chromatography (MEKC) to obtain protein expression fingerprints. Analytes were detected in a sheath flow cuvette using laser-induced fluorescence. In a 1-h 2D-CE separation, over 100 components were resolved with a spot capacity of 380.
Keywords: Single zygote analysis; Two-dimensional capillary electrophoresis; Single cell analysis
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