A long-wavelength biolabeling reagent based on the oxonol fluorophore |
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Authors: | Philip L. Southwick Klaus M. Hahn Jean Chao Patrick L. Perry Allan S. Wagman Marc Wagner Alan S. Waggoner |
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Affiliation: | (1) Center for Light Microscope Imaging and Biotechnology, Carnegie Mellon University, 4400 5th Avenue, 15213 Pittsburgh, Pennsylvania;(2) Division of Virology, Department of Neuropharmacology, Scripps Research Institute, La Jolla, California |
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Abstract: | A red fluorescent dye of the oxonol class, bis-[1-(carboxymethyl)barbituric acid-(5)]-pentamethinoxonol, has been synthesized and, in the form of the succinimidyl active ester, has been applied to antibody labeling for application to flow cytometry and to imaging of tissue sections. The new dye, named CMOX (for carboxymethyloxonol), shows maximum excitation at 583 nm and emission at 611 nm, with a quantum yield of 0.2 in aqueous buffer and methanol. Antibodies labeled with the new dye show favorable brightness, photostability, and low levels of nonspecific binding. |
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Keywords: | Oxonol succinimidyl ester fluorescent labeling flow cytometry immunofluorescence |
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