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- W166231307 abstract "Establishment of an accurate extinction coefficient is often of interest when a precise amount of material is required for a biochemical reaction or when converting an association constant from absorbance units into concentration units. Although a gravimetric approach can provide a quick estimate of the concentration, the presence of salts or impurities can adversely affect its accuracy. Analytical ultracentrifugation is often used when more accurate estimates of the extinction coefficient are required. This technique has been applied to prostate specific antigen (PSA), and the value of the extinction coefficient estimated by amino acid analysis is confirmed by sedimentation analysis.The extinction coefficient of PSA was measured by analytical ultra-centrifugation using the method of sedimentation boundary analysis. This method utilizes centrifugal force to form a solvent boundary by layering a reference buffer onto a sample via a thin capillary connecting both sectors of the centerpiece i.e., a synthetic boundary centerpiece. The plateau formed by the boundary is then optically measured as it decreases over time due to diffusional effects. By using a combination of absorbance and interference optics, the OD and concentration (from the fringe displacement) can be sequentially measured and used in Beer’s Law to directly solve for the extinction coefficient. A value for the extinction coefficient of PSA at 280 nm of 1.71 L/g-cm was obtained. This is in close agreement with a literature value of 1.84 L/g-cm using a concentration estimated by amino acid analysis." @default.
- W166231307 created "2016-06-24" @default.
- W166231307 creator A5047461059 @default.
- W166231307 date "2007-12-04" @default.
- W166231307 modified "2023-10-17" @default.
- W166231307 title "Measurement of the extinction coefficient of prostate specific antigen using interference and absorbance optics in the Optima XL-A analytical ultracentrifuge" @default.
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- W166231307 doi "https://doi.org/10.1007/bfb0114086" @default.
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