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Allophycocyanin is a pigment from the Phycobiliprotein family. It is a bluish green protein found in Cyanobacteria species Spirulina Plantensis, with high fluorescence quantum yield, molar extinction coefficients, absorbtivity and stability. It can be easily cross linked with antibodies and other proteins by using conventional molecular tagging techniques without losing its fluorescent properties. It is composed of α and β subunits and these subunits are present in trimer form. Each subunit has a molecular weight of 17 K Dalton. The total molecular weight of Allophycocyanin is 104 K Dalton.
Allophycocyanin has absorption maximum at 650 nm and emission maximum at 660 nm; it is an ultra sensitive florescent tracer because of high emission quantum yield and is 100 times more sensitive than conventional organic fluorophores. It has therapeutic properties and is also used as food colorant having an isoelectric point of 4.36, trimer is unstable at low concentration and trimer has got higher emission quantum yield than monomer.
High Extinction Coefficients and Quantum Yield: Allophycocyanin based dyes exhibit very intense fluorescence light emission and optimal brightness due to high extinction coefficients and quantum yield values of the dyes - intensity increases when coupled to bio molecules resulting in reduced influence of uncoupled dye.
High Resistance to Photo Bleaching: As compared to other fluorescent dyes phycobiliprotein based dyes can withstand higher photo-degradation, hence they show lesser photo bleaching effect.
pH Stability: Phycobiliprotein based dyes are stable at a wider range of pH as compared to other fluorescent dyes( pH range- 5 to 8).
Hydrophilic in Nature: The negatively charged Allophycocyanin based dyes are highly water soluble and thus can be used in biological test done in vitro and in vivo.
High water solubility.
Good instrument capability.
Stability over broad pH range.
High affinity towards genomic DNA.
Free from interference from auto fluorescence of cellular components.
1 FRET Assays
2 Flow Cytometry
3 Fluorescence Activated Cell Sorting (FACS).
4 Fluorescence in Situ Hybridization (FISH).
5 Labeling of proteins, antibodies and nucleic acids.
6 Fluorescence microscopy
7 Bio Maker: Allophycocyanin has a great utility in biomarkers because of its high fluorescence, but typically in combination with R-PE to form a fluorescent tandem phycobiliprotein conjugate which improves sensitivity.
8 As a Pharmacologically Active Compound: Allophycocyanin is applied in health care products due to certain functional effects such as inhibition of enterovirus 71- induced apoptosis. At concentration nontoxic to the host cells (0.045±0.012 μM), allophycocyanin was found to inhibit enterovirus 71-induced cytopathic effects, viral plaque formation, and viral-induced apoptosis.
9 As Food Colorant: It used as a food colorant because of its algal based source, hydrophilic nature and stability at room temperature.
1 A651/A280 >4.0
2 A651/A280 >3.0
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