Isoelectric focusing in layers of granulated gels. II. Preparative isoelectric focusing.

摘要: Abstract A method for preparative isoelectric focusing of 0.1–10 g amounts proteins is described. For anticonvective stabilization the pH gradient, layers granulated gels (e.g. Sephadex or Bio-Gel) variable length, width and thickness were used either on glass plates in troughs. Load capacity, defined as amount protein per ml gel suspension, was determined to be 5–10 mg total protein, irrespective range carrier ampholytes. single load capacities 0.25–1 found 3–10 ampholytes, 2–4 narrow Experiments a quartz plate followed by densitometric evaluation situ at 280 nm have demonstrated that it possible proceed from analytical thin-layer separations without loss resolution, just changing dimension layer increasing load. Improved resolution which facilitates isolation isoelectrically homogeneous components could achieved 40 cm long separation distance. The geometry favourable heat dissipation thus permits use high voltage gradients. Recovery focused elution simple. efficiency illustrated examples showing mixtures.