By John N. Abelson(Editor), Melvin I. Simon(Editor), Harry Walter(Editor), Gote Johansson(Editor)
This quantity of tools in Enzymology represents the only real choice of separation tools in keeping with aqueous two-phase structures. It comprises systems for the isolation of proteins, specially enzymes, nucleic acids, cellphone membranes, and organelles, in addition to for the separation and examine of cells. Key positive factors* Use of affinity partitioning for enzyme purification* Separation and learn of cells* Isolation of plasma membranes* huge scale biotechnical use
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Cell Res. ] relative electrophoretic mobilities. The exceptions to the correlation indicate that the surface charge measured by electrophoresis and partitioning can differ and that surface properties other than charge can also affect the partition ratio obtained (see Ref. 1 for detailed discussion). Non-Charge-Sensitioe Phase Systems. , in systems closer to the critical point), the interfacial tension diminishes and the partition ratios increase. Even in phases containing NaCI measurable partition ratios often result.
Systems with polymer concentrations such as those used in the noncharge-sensitive phases described above into which phosphate, for example, is incorporated will have both charge-sensitive and non-charge-sensitive characteristics. Charge-sensitive systems into which a polymerligand is incorporated may also reflect those properties. Methods Preparation of Particulate Material for Partitioning Cells. As indicated above the partitioning behavior of cells depends on their surface properties. If one is interested primarily in effecting a cell separation, and not in the original surface properties of the cells, the preparation of single cells by any means (enzymatic, mechanical) that does not alter the biological property of interest can be used.
Kopperschl~iger, this volume [11]. 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 [3] 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. Phase system: dextran TS00, PEG 8000, and 50 mM K2SO4. Temperature, 20°. [Based on data from G. Johansson, J. Chromatogr. ] (B) Variation of K~, for phosphofructokinase from yeast using Cibacron Blue F3G-A-PEG, with increasing s values.