Comparison of DIGE and post-stained gel electrophoresis with both traditional and SameSpots analysis for quantitative proteomics.
2-DE is an important tool in quantitative proteomics. Here, we compare the deep purple (DP) system with DIGE using both a traditional and the SameSpots approach to gel analysis. Missing values in the traditional approach were found to be a significant issue for both systems. SameSpots attempts to address the missing value problem. SameSpots was found to increase the proportion of low volume data for DP but not for DIGE. For all the analysis methods applied in this study, the assumptions of parametric tests were met. Analysis of the same images gave significantly lower noise with SameSpots (over traditional) for DP, but no difference for DIGE. We propose that SameSpots gave lower noise with DP due to the stabilisation of the spot area by the common spot outline, but this was not seen with DIGE due to the co-detection process which stabilises the area selected. For studies where measurement of small abundance changes is required, a cost-benefit analysis highlights that DIGE was significantly cheaper regardless of the analysis methods. For studies analysing large changes, DP with SameSpots could be an effective alternative to DIGE but this will be dependent on the biological noise of the system under investigation.
Karp
Natasha A
NA
Department of Biochemistry, Cambridge University, Cambridge, England.
Feret
Renata
R
Rubtsov
Denis V
DV
Lilley
Kathryn S
KS
D000818:Animals; D015180:Electrophoresis, Gel, Two-Dimensional / Q000191:economics / Q000379:methods*; D005453:Fluorescence; D051379:Mice; D016973:Pectobacterium carotovorum / Q000737:chemistry; D040901:Proteomics