Download PDF by G. D. Birnie, D. Rickwood: Centrifugal Separations in Molecular and Cell Biology

Download PDF by G. D. Birnie, D. Rickwood: Centrifugal Separations in Molecular and Cell Biology

By G. D. Birnie, D. Rickwood

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In general, however, the best approach is to unload the whole gradient into a number of fractions whenever this is possible, even if the particles to be recovered occupy only a single, narrow zone. 7. Note particularly that the gradient is not inverted at any point (except just above the fraction tubes) and that the gradient is 'funnelled' out of the tube and into and out of the photometer cell. Unloading systems such as this have been investigated by Brakke (1963) and others, who found that this form of unloading gave better results than direct withdrawal of a zone through the side of the tube with a syringe needle.

G. (1955). Expl Cell. , 9, 446 (1966a). Natn, Cancer Inst. Monogr,, No. 21, p . 9 (19661)). L. M. (1963). In Ultracentrifugal Analysis, p. 143. Ed. T. Williams. TVflm. Cancer Inst. , No. 2, p . S. (1966). TVfl/w. Cancer Inst. , No. 2 1 , p . , RICKWOOD, D. and HELL, A. (1973). Biochim biophys. Acta, 331, 283 BISHOP, B. {1966), Natn. , No. 2 1 , p . K. /. Am. K. {1964). ArchsBiochem. F. and EISENBERG, Η. (1964). Adv. , G I R A U D , B. and MESSIAH, A. (1967). J. {1966). J. ArchsBiochem. H. and HORTON, j .

One important point that should be considered is the rates of acceleration and decelerarion of rotors at the beginning and end of runs. It has been found that sudden changes in the angular velocity, especially below 5000 rev/min, can cause a 'swirling' motion of the gradient solurion. This effect is particularly pro­ nounced when non-viscous gradients and/or wide-diameter tubes are employed. 2 Centrifugation CENTRIFUGATION CONDITIONS: EXAMPLES OF TIME, FORCE A N D GRADIENT FOR VARIOUS RATE-ZONAL SEPARATIONS.

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