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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
biochemistry, genetics and molecular biology
The use of 3-aminophthalimide as a pro-chemiluminescent label in chemiluminescence and fluorescence-based cellular assays
Bioconjugate Chemistry, Volume 20, No. 2, Year 2009
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Description
We present a labeling system for direct chemiluminescence-based cellular bioassays using the stable prochemiluminescent, luminol precursor, 3-aminophthalimide (API). API-coupled reporter molecules are detected chemiluminometrically after treatment with hydrazine, which converts the API label to luminol. API derivatives containing a variety of functional groups are readily synthesized, allowing for ease of coupling via the imide nitrogen to a host of reporter molecules. The fluorescent nature of APIs further allows for dual fluorescence and chemiluminescence studies. To highlight the utility of this label, we show that API-labeled insulin can be successfully utilized in cellular binding and transport assays and that an API-coupled mitochondrial probe (API-triphenylphosphonium+) can be used to both fluorescently and chemiluminometrically investigate mitochondrial function. We also assess the use of API as a polysaccharide and nucleic acid label, and we show that API-labeled palmitic acid undergoes cellular transport and lipid metabolism. © 2009 American Chemical Society.
Authors & Co-Authors
Mavri-Damelin, Demetra
United Kingdom, London
Ucl Medical School
South Africa, Johannesburg
University of the Witwatersrand
Wilden, Jonathan
United Kingdom, London
University College London
Mani, Ali Reza
United Kingdom, London
Ucl Medical School
Selden, Clare A.
United Kingdom, London
Ucl Medical School
Hodgson, Humphrey J.F.
United Kingdom, London
Ucl Medical School
Damelin, Leonard Howard
United Kingdom, London
Ucl Medical School
South Africa, Johannesburg
National Institute for Communicable Diseases
Statistics
Citations: 6
Authors: 6
Affiliations: 4
Identifiers
Doi:
10.1021/bc800361r
ISSN:
10431802