Impact of quenching failure of Cy dyes in differential gel electrophoresis.

ABSTRACT:

Background

Differential gel electrophoresis (DIGE) is a technology widely used for protein expression analysis. It is based on labelling with fluorescent Cy dyes. In comparative fluorescence gel electrophoresis experiments, however, unspecific labelling using N-hydroxy-succinimide-ester-based labelling protocols was recently detected. Cross-talk was observed due to failure of the quenching process. Here, the impact of this effect for DIGE experiments was investigated.

Methodology/principal findings

Experiments to test quenching efficiency were performed in replicate using Escherichia coli lysate. Parameters such as the amount of dye and quencher were varied. Labelling and quenching were reversed in one experiment. Differences in protein spot volumes due to limited quenching

SUBMITTER: Wang W 

PROVIDER: S-EPMC3068157 | biostudies-literature | 2011 Mar

REPOSITORIES: biostudies-literature