Cryptomonad algal phycobiliproteins as fluorochromes for extracellular and intracellular antigen detection by flow cytometry

Cytometry
W G TelfordF C Allnutt

Abstract

Phycobiliproteins play an important role in fluorescent labeling, particularly for flow cytometry. The spectral properties of R-phycoerythrin (R-PE) and allophycocyanin (APC) have made them the dominant reagents in this class of fluorochromes. In this study, we evaluate a lesser-known but potentially important series of low-molecular weight cryptomonad-derived phycobiliproteins (commercially termed the CryptoFluortrade mark dyes) for their applicability to flow cytometry, both in extracellular and intracellular labeling applications. Several cell lines were labeled with biotin-conjugated antibodies against expressed extracellular surface proteins, followed by streptavidin conjugates of three cryptomonad phycobiliproteins (CryptoFluor-2, CryptoFluor-4, and CryptoFluor-5). Cells were then analyzed by flow cytometry using a variety of laser lines and emission filters to establish the optimal excitation/emission characteristics for each fluorochrome. Some cells were permeabilized and labeled for intracellular antigens, also using the cryptomonad fluorochromes. Where appropriate, parallel samples were labeled with other fluorochromes (including R-PE, APC, the cyanin dyes Cy3 and Cy5, and others) to gauge the performance of the crypt...Continue Reading

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Citations

Apr 15, 2010·Avian Pathology : Journal of the W.V.P.A·S De BoeverE Meyer
Sep 26, 2013·BioMed Research International·M KuddusAwdah Al-Hazimi
Feb 13, 2015·Critical Reviews in Food Science and Nutrition·Hasina BegumMohamed Shariff
Nov 19, 2003·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·Judith E BerlierRichard P Haugland
Mar 17, 2016·World Journal of Biological Chemistry·Ravi Raghav SonaniDatta Madamwar

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