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  • Gel-based separation of phosphoproteins in samples stored in urea/thiourea after precipitation by lanthanum chloride.

Gel-based separation of phosphoproteins in samples stored in urea/thiourea after precipitation by lanthanum chloride.

Electrophoresis (2012-11-30)
Mario Pink, Christine Stein, Nisha Verma, Albert W Rettenmeier, Simone Schmitz-Spanke
要旨

The recent introduction of the La(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 2DE.

材料
製品番号
ブランド
製品内容

Sigma-Aldrich
塩化ランタン(III) 七水和物, ACS reagent
Sigma-Aldrich
塩化ランタン(III) 七水和物, 99.999% trace metals basis
Sigma-Aldrich
塩化ランタン(III), AnhydroBeads, −10 mesh, ≥99.99% trace metals basis
Sigma-Aldrich
塩化ランタン(III), AnhydroBeads, −10 mesh, 99.9% trace metals basis