Method of determining extinction coefficient of fluorescent dye and protein concentration of dye-protein conjugate
US5245551A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | Apr 10, 1991 |
| Grant date | Sep 14, 1993 |
| Priority date | — |
| Expiry date | Apr 10, 2011 |
Classification
- Technology area (CPC G)Physics
- CPC primaryG01N2201/129
- WIPO fieldMeasurement
- WIPO sectorInstruments
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: EQU A.sub.p-d =.alpha..multidot.A.sub.p +.beta..multidot.A.sub.d +.epsilon. where, A.sub.p-d, A.sub.p, A.sub.d are the absorption spectra of the dye-protein conjugate, the protein alone, and the dye-polypeptide compound, respectively, .alpha., .beta. are the regression coefficients to be determined, and .epsilon. is the error term.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.