In conventional isoelectric focusing in soluble buffers, it has been quite difficult to produce two-dimensional (2-D) separation sin pH intervals greater than pH 4-8. In general more alkaline proteins wer analyzed by non-equilibrium IEF in the first dimension. Even with the advent of immobilized pH gradients (IPG), separations could be extended to pH gradients not wider than pH 3-10, due to a lack of suitable buffers. Since more acidic and more alkaline acrylamido buffers have recently been synthesized, we have been able to optimize what is beleived to be the widest possible immobilized pH gradient, a h 2.5-11 span. We report here for the first time 2-D separations of total tissue lysates in such extended pH 2.5-11 gradients. It appears that, with the IPG technique, close to 100% of all possible cell products can be displayed in a single 2-D map.

2-DIMENSIONAL MAPS IN THE MOST EXTENDED (PH 2.5-11) IMMOBILIZED PH GRADIENT INTERVAL / P. SINHA, M. PRAUS, E. KOTTGEN, E. GIANAZZA, P. RIGHETTI. - In: JOURNAL OF BIOCHEMICAL AND BIOPHYSICAL METHODS. - ISSN 0165-022X. - 21:2(1990), pp. 173-179.

2-DIMENSIONAL MAPS IN THE MOST EXTENDED (PH 2.5-11) IMMOBILIZED PH GRADIENT INTERVAL

E. GIANAZZA
Penultimo
;
1990

Abstract

In conventional isoelectric focusing in soluble buffers, it has been quite difficult to produce two-dimensional (2-D) separation sin pH intervals greater than pH 4-8. In general more alkaline proteins wer analyzed by non-equilibrium IEF in the first dimension. Even with the advent of immobilized pH gradients (IPG), separations could be extended to pH gradients not wider than pH 3-10, due to a lack of suitable buffers. Since more acidic and more alkaline acrylamido buffers have recently been synthesized, we have been able to optimize what is beleived to be the widest possible immobilized pH gradient, a h 2.5-11 span. We report here for the first time 2-D separations of total tissue lysates in such extended pH 2.5-11 gradients. It appears that, with the IPG technique, close to 100% of all possible cell products can be displayed in a single 2-D map.
Immobilized pH gradient; Isoelectric focusing; Two-dimensional map
Settore BIO/10 - Biochimica
1990
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/180889
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