Jump to main content
Jump to site search

Issue 19, 2017
Previous Article Next Article

Quantification of green fluorescent protein-(GFP-) tagged membrane proteins by capillary gel electrophoresis

Author affiliations

Abstract

A fast and robust procedure for the quantification of GFP-tagged membrane proteins in cell homogenates was developed employing capillary gel electrophoresis coupled to laser-induced fluorescence detection (CGE-LIF). The new method was found to be highly sensitive and applicable to structurally diverse membrane proteins including synaptic vesicle protein 2A (SV2A), adenosine A2A receptor (A2AAR), and connexin 43 (Cx43). Quantification of SV2A and A2AAR using radioligand binding assays confirmed the results obtained with CGE-LIF. The CGE-LIF method showed significantly higher sensitivity as compared to fluorimetric measurement in a microplate. Importantly, CGE-LIF involves separation of the target proteins and their degradation products prior to quantification and thereby ensures specificity. We anticipate broad applicability of the method for any fluorophore-tagged protein.

Graphical abstract: Quantification of green fluorescent protein-(GFP-) tagged membrane proteins by capillary gel electrophoresis

Back to tab navigation

Supplementary files

Publication details

The article was received on 13 Jun 2017, accepted on 22 Aug 2017 and first published on 23 Aug 2017


Article type: Paper
DOI: 10.1039/C7AN00981J
Citation: Analyst, 2017,142, 3648-3655
  •   Request permissions

    Quantification of green fluorescent protein-(GFP-) tagged membrane proteins by capillary gel electrophoresis

    A. Danish, S. Lee and C. E. Müller, Analyst, 2017, 142, 3648
    DOI: 10.1039/C7AN00981J

Search articles by author

Spotlight

Advertisements