In mixed-bed, carrier ampholyte-Immobiline gels, a primary, insolubilized pH gradient is admixed with a secondary, soluble pH gradient generated by amphoteric buffers. The latter are the standard carrier ampholytes (e.g. Ampholine, Pharmalyte, Biolyte, Servalyte), used in conventional isoelectric focusing, admixed to Immobiline gels in levels of approximately 0.5-1%. It is here shown that polybuffers 96 (covering the pH 6-9 range) and 74 (covering the pH 4-7 interval) used as eluents in chromatofocusing, can effectively substitute the standard carrier ampholytes with considerable savings (they are 1/16th as expensive as the latter chemicals).