By John N. Abelson(Editor), Melvin I. Simon(Editor), Harry Walter(Editor), Gote Johansson(Editor)
This quantity of tools in Enzymology represents the only choice of separation tools in accordance with aqueous two-phase structures. It comprises techniques for the isolation of proteins, particularly enzymes, nucleic acids, cellphone membranes, and organelles, in addition to for the separation and learn of cells. Key positive aspects* Use of affinity partitioning for enzyme purification* Separation and research of cells* Isolation of plasma membranes* huge scale biotechnical use
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Additional info for Aqueous Two-Phase Systems
2. K0 value for bovine serum albumin, partitioned at its isoelectric point, as a function o f tie-line length, s, in systems containing dextran T500 and PEG 8000 (©) or P E G 35,000 (0). Temperature, 20 °. [Reproduced from G. Johansson, J. Chromatogr. ] T A B L E II SPECIFIC VALUES, 3'+ AND 3 ' _ , FOR CATIONS AND ANIONS FOR CALCULATION OF RELATIVE 3' VALUES a'b Cation K+ Na + NH4 + Li + (C2H5)4N+ (C4Hg)4N + y+ Anion 3'_ 58 56 49 43 32 6 H2PO4-/HPO42- (1 : 1) SO42FAcetate CIBrISCNClO 4- 79 77 69 56 50 44 34 30 28 a Obtained with different salts in Dextran T500-poly(ethylene glycol) 8000 phase systems at 0 - 4 °.
A. Albertsson, Eur. J. Biochem. 33, 379 (1973). 20 L. Cheng, M. Joelsson, and G. Johansson, J. Chromatogr. 523, 119 (1990). 21 G. Johansson, this volume . 22 G. Johansson, J. Chromatogr. 451, 517 (1976). 23 G. Kopperschl~iger, this volume . 24 S. D. Flanagan and S. H. Barondes, J. Biol. Chem. 2,50, 1484 (1975). 25 G. Johansson and F. Tjerneld, J. Biotechnol. U , 135 (1989). 34 GENERAL METHODOLOGY AND APPARATUS  A 0 t I J 20 10 0 $ B 3 3:: ~z 00 = I = 10 I 20 S FIG. 3. (A) Variation of Khp, for bovine serum albumin using PEG-palmitate, with increasing tie-line length, s.
Increased cell quantities generally require a corresponding increase in ligand concentration to effect adequate extraction. ORGANELLES AND MEMBRANES. With organelles and membranes the usual goal is to separate one particular kind from all the others in a complex mixture rather than to compare surface properties. Relatively rapid preparative procedures are desired so as to better preserve biological activities. The best scenario is when the material of interest partitions into either the top phase or to the interface plus bottom phase and the rest of the [4l PARTITIONING PROCEDURES: PARTICULATES 55 material to the opposite phase.