Method of determining extinction coefficient of fluorescent dye

Details Method of determining extinction coefficient of fluorescent dye Method of determining extinction coefficient of fluorescent dye

1991-04-10

1993-09-14

Cosimano, Edward R.

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364498, G06F 1520

Patent

active

052455513

ABSTRACT:
The extinction coefficient of fluorescent dyes in donor-acceptor energy transfer processes is determined for dye-protein conjugates by forming short polypeptide fragment-dye conjugates and measuring the change in ultraviolet light absorbance of the conjugates with changing concentration. The protein concentration is determined by comparing the absorption spectrum of a conjugate of donor dye, acceptor dye, and protein with the individual components thereof using a multiple linear regression technique based on the following model:

REFERENCES:
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patent: 5051249 (1991-09-01), Metcoff
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Neter et al: "Applied Linear Regression Models"; Irwin Inc.; ISBN 0-256-07068-7; 1989 pp. xiii, 225-236.
Beebe et al: "An Introduction to Multivariate Calibration and Analysis"; Analytical Chemistry, vol. 59 No. 17, Sep. 1, 1987; pp. 1007A-1017A.
Gemperline et al: "Background Correction in Multicomponent Spectroscopic Analysis Using Target Transformation Factor Analysis"; Applied Spectroscopy; vol. 41, No. 3; 1987 pp. 454-459.
I. C. McKay et al., "A comparison of Fluorescein Isothiocyanate and Liassamine Rhodamine (RB 200) As Labels for Antibody in the Fluorescent Antibody Technique", Immunology 1981, 43, 591-602.
P. L. Khanna, "Fluorescence Energy Transfer Immunoassays", Plenum Press, 1988, pp. 211-229.

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