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Determination of phycobiliproteins by capillary electrophoresis with laser-induced fluorescence detection

✍ Scribed by Pertti J. Viskari; Christopher S. Kinkade; Christa L. Colyer


Publisher
John Wiley and Sons
Year
2001
Tongue
English
Weight
118 KB
Volume
22
Category
Article
ISSN
0173-0835

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✦ Synopsis


Determination of phycobiliproteins by capillary electrophoresis with laser-induced fluorescence detection

Phycobiliproteins are derived from the photosynthetic apparatus of cyanobacteria and eukaryotic algae. They are composed of a protein backbone to which linear tetrapyrrole chromophores are covalently bound. Furthermore, they are water-soluble, highly fluorescent, and relatively stable at room temperature and neutral pH. For this reason, capillary electrophoresis-laser induced fluorescence (CE-LIF) seems the ideal method for determination of these important proteins. The effects of buffer additives such as sodium dodecyl sulfate (SDS)and putrescine on the separation of the three major phycobiliprotein types, namely allophycocyanin, phycocyanin, and phycoerythrin, with excitation and emission maxima at 652/660, 615/647, and 565(494)/575 nm, respectively, are considered. Detection limits for these proteins by CE-LIF are some 60-500 times better than by absorbance detection. The development of a fast and sensitive CE-LIF assay such as this is of potential significance to our understanding of chemical and biological oceanographic processes.


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Aequorea victoria green fluorescent protein was assayed by capillary electrophoresis using post-capillary laserinduced fluorescence detection in a sheath flow cuvette. The limit of detection was 3.0 Ο« 10 Οͺ 12 M protein in an injection volume of 17 nL, corresponding to a mass of 3100 molecules.