Isoelectric focusing immobilized

Fig. 1. 2D-PAGE. 180. ig of platelet proteins Is separated. For isoelectric focusing, immobilized pH gradient strips (pH 3-10) have been used.

Righetti, P. G., Isoelectric focusing of proteins in conventional and immobilized pH gradients, in Protein Structure A Practical Approach, Creighton, T. E., Ed., IRL Press, New York, 1989, 23. 

Ek, K., Bjellqvist, B., Righetti, R G. (1983). preparative isoelectric-focusing in immobilized pH Gradients. 1. General principles and methodology. J. Biochem. Bioph. Meth. 8(2), 135-155. 

Essader, A.S., Cargile, B.J., Bundy, J.L., Stephenson, J.L., Jr. (2005). A comparison of immobilized pH gradient isoelectric focusing and strong-cation-exchange chromatography as a first dimension in shotgun proteomics. Proteomics 5, 24—34. 

Norbeck J et al. Two-dimensional electrophoretic separation of yeast proteins using a non-linear wide range (pH 3-10) immobilized pH gradient in the first dimension reproducibility and evidence for isoelectric focusing of alkaline (pi >7) proteins. Yeast 1997 13 1519-1534. 

PI. Pirttila, T., Frey, H., Mattila, K., and Siden, L., Immobilized pH gradient isoelectric focusing of cerebrospinal fluid proteins. Electrophoresis 9, 582-588 (1988). 

Haase R, Menke-Mollers I, Oette (1988) Analysis of human apolipoproteins C by isoelectric focusing in immobilized pH gradients. Electrophoresis 9 569-575... 

Fig. 17.6. Isoelectric focusing diagrammatic view of isoelectric focusing (IEF) of a re-hydrated linear immobilized pH 3-10 gradient strip (Biorad, UK).

PG Righetti, A Bossi. Isoelectric focusing in immobilized pH gradients an update. J Chromatogr B 699 77-89, 1997. 

Figure 8.8. Resolution by isoelectric focusing in an immobilized pH gradient (pH range 7.35-7.55 over 20 cm path) of two human fetal hemoglobins, one with glycine (lower) instead of alanine (upper) in position 136 of the gamma chains. The isoelectric points of the hemoglobins differ by only 0.003 pH units. (Courtesy of P. G. Righetti, University of Milano.)...

Righetti, P. G., and Bossi, A. (1997). Isoelectric focusing in immobilized pH gradients Recent analytical and preparative developments. Anal. Biochem. 247, 1-10. 

Stoyanov, A. V., and Righetti, P. G. (1998). Steady-state concentration distribution of ampholytes in isoelectric focusing in a linear immobilized pH gradient. Electrophoresis 19, 1596-1600. 

Wenisch, E., Jungbauer, A., Tauer, C., Reiter, M., Gruber, G., Steindl, F., and Katinger, H. (1989). Isolation of human monoclonal antibody isoproteins by preparative isoelectric focusing in immobilized pH gradients. J. Biochem. Biophys. Methods 18, 309-322. 

One of the most troublesome aspects of isoelectric focusing is how to modulate the slope of the pH gradient to increase resolution (equivalent to pH gradient engineering, as easily available in immobilized pH gradients). A simple solution is offered with addition, to the standard 2-pH-units interval, of separa-... 

Westeemeiee, R., Postel,W. (1982). Isoelectric focusing in immobilized pH gradients principle, methodology and some applications. /. Biochem. Biophys. Methods 6(4), 317-339. 

Gorg, A., PosTEL, W., Gunther, S., Weser, J. (1985). Improved horizontal two-dimensional electrophoresis with hybrid isoelectric focusing in immobilized pH gradients in the first dimension and laying-on transfer to the second dimension. Electrophoresis 6, 599-604. 

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