William B. Jakoby (Eds.)'s Enzyme purification and related techniques PDF

By William B. Jakoby (Eds.)

ISBN-10: 0121818853

ISBN-13: 9780121818852

The significantly acclaimed laboratory regular, Methods in Enzymology, is likely one of the so much hugely revered courses within the box of biochemistry. considering the fact that 1955, each one quantity has been eagerly awaited, often consulted, and praised by means of researchers and reviewers alike. The sequence includes a lot fabric nonetheless correct this present day - really a necessary book for researchers in all fields of lifestyles sciences

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T h e crank is rotated by a motor, at 50 r p m , to p r o d u c e a wagging motion o f the cell. Using such an apparatus, 9 6 % o f the salt was r e m o v e d f r o m 1 ml o f a 2 M a m m o n i u m sulfate solution in 30 minutes. In n o r m a l s u s p e n d e d dialysis, only 6 0 % o f the salt was r e m o v e d in an equal period o f time. FIG. 3. The rapid dialyzer of E. Cabib and I. D. Algranati. Reproduced from Nature (London) [ 188, 409 ( 1960)] by courtesy of the authors and publishers. Cabib and I.

9 M ammonium sulfate solution were dialyzed in this apparatus through 18/32 tubing and against distilled water, 99% of'the salt was removed in 7 hours. The total volume of diffusate was 650 ml. Lauffer has described a method by which the rocking dialyzer can be converted into a circulatory systemY By including two simple check valves in the circuit, the rocking motion is used to drive diffusate through the cylinder, from an external reservoir. The Dialysis of Large V o l u m e s - Countercurrent Dialysis Obviously, when the volume of retentate to be treated becomes much larger than 100 ml, the lengths of dialysis tubing and the volumes of diffusate which must be used in any of the above techniques soon become relatively impractical.

TM A glass plug is formed by sealing the ends of a piece of 2-cm diameter glass tubing. The length of the plug is determined by the volume of solution to be dialyzed. For a 1-ml sample, the plug should be 2 cm in length. Before the plug is finally sealed, sufficient mercury is introduced into it so that it will barely float. The plug is slipped into a length of size 27 dialysis tubing, which is sealed just below the plug. The sample is introduced with a Pasteur pipette and the tubing sealed just above the plug.

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Enzyme purification and related techniques by William B. Jakoby (Eds.)


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