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Gel‐based separation of phosphoproteins in samples stored in urea/thiourea after precipitation by lanthanum chloride
Author(s) -
Pink Mario,
Stein Christine,
Verma Nisha,
Rettenmeier Albert W.,
SchmitzSpanke Simone
Publication year - 2013
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201200278
Subject(s) - urea , lysis , thiourea , phosphoprotein , lysis buffer , chromatography , precipitation , chemistry , lanthanum , biochemistry , inorganic chemistry , phosphorylation , organic chemistry , physics , meteorology
The recent introduction of the L a 3+ precipitation method for the enrichment of phosphoproteins allows a gel‐based analysis of these posttranslationally modified proteins. However, if this method is applied to cell lysates stored in urea‐containing lysis buffer for an extended period of time, incomplete phosphoprotein recovery is observed. We ascribe this effect to the presence of urea in the lysis buffer. To overcome this problem various strategies were tested, where cell lysates stored at least for one year were utilized. By applying an optimized protocol approximately 250 proteins could be observed following separation by 2 DE .