By Alfred W. Francis
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ACS Symposium Series; American Chemical Society: Washington, DC, 1987. Chapter 3 Immobilized pH Gradients: Recent Developments Pier Giorgio Righetti, Cecilia Gelfi, and Elisabetta Gianazza University of Milano, Via Celoria 2, Milano 20133, Italy The present revie isoelectric focusing in immobilized pH gradients in three main areas: a) "mixed bed" Ampholine-Immobiline gels; b) two dimensional maps and c) preparative runs. Topic (a) presents the advantages of using a combination of a primary, immobilized gradient whith a superimposed secondary, carrier ampholyte pH gradient.
15. 16. 17. 18. 19. 20. 21. 22. 23. 24. 25. 26. 27. 28. 29. 30. 31. 32. 33. 34. 35. 36. 37. 38. G. "Isoelectric focusing: theory, methodology and applications"; Elsevier: Amsterdam, 1983. R. ; "Gel Electro phoresis and Isoelectric Focusing of Proteins: Selected Techniques"; de Gruyter: Berlin, 1984. Neuhoff, V. : "Electrophoresis '84", Verlag Chemie: Weinheim, 1984 Chrambach, A. "The Practice of Quantitative Gel Electrophoresis"; VCH Verlagsgesellschaft: Weinheim, 1985. J. : "Gel Electrophoresis of Proteins"; Wright: Bristol, 1986.
While i t appears that most membrane proteins have about the same o v e r a l l hydrophobicity as ordinary water-soluble proteins, i t cannot be excluded that t h e i r hydrophobic amino acids could be concentrated i n c e r t a i n regions of the molecule, or be more exposed on the surface, thus imparting to them a strong tendency to aggregate once i n s o l u t i o n . The presence of CAs could be b e n e f i c i a l t o t h e i r s o l u b i l i t y both, i n the t r a n s i e n t and steady-state, as i t i s known that, e s p e c i a l l y when focused i n the p i zone, t h e i r backbone e x h i b i t s more hydrophobic c h a r a c t e r i s t i c s , as most of the i o n i z a b l e groups tend t o be i n the uncharged state (36).