Method for Pre-Fractionation of Complex Samples
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United States of America Patent
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Issued Date -
Dec 25, 2008
app pub date -
Nov 14, 2006
filing date -
Nov 18, 2005
priority date (Note) -
Abandoned
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Abstract
The present invention relates to a method for pre-fractionation of complex protein and/or peptide samples. The method comprises the following steps: a) loading the sample onto pre-packed media able to separate proteins/peptides according to their pl-value; b) eluting the media by centrifugal force under denaturing conditions with at least three buffers having different pH in a stepwise manner to obtain at least three fractions separated according to pl-value; and c) subjecting each fraction to further separation in a pH-range corresponding to the pl-values of said fractions. The invention also relates to a kit comprising at least one spin column or multiwell plate filled with chromatography medium able to separate proteins/peptides according to pl-value, at least three buffers, and one or more IPG strips having a narrow pH-range adapted to each of said buffers.
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