论文信息 - Evaluation of individual protein errors in silver-stained two-dimensional gels.

Evaluation of individual protein errors in silver-stained two-dimensional gels.

The relationship between spot volume and variation for all protein spots observed on large format 2D gels when utilising silver stain technology and a model system based on mammalian NSO cell extracts is reported. By running multiple gels we have shown that the reproducibility of data generated in this way is dependent on individual protein spot volumes, which in turn are directly correlated with the coefficient of variation. The coefficients of variation across all observed protein spots were highest for low abundant proteins which are the primary contributors to process error, and lowest for more abundant proteins. Using the relationship between spot volume and coefficient of variation we show it is necessary to calculate variation for individual protein spot volumes. The inherent limitations of silver staining therefore mean that errors in individual protein spot volumes must be considered when assessing significant changes in protein spot volume and not global error.

[1] J. Nishihara,et al. Quantitative evaluation of proteins in one‐ and two‐dimensional polyacrylamide gels using a fluorescent stain , 2002, Electrophoresis.

[2] M. Wilkins,et al. Progress with gene‐product mapping of the Mollicutes: Mycoplasma genitalium , 1995, Electrophoresis.

[3] P Dupree,et al. Quantitative and reproducible two‐dimensional gel analysis using Phoretix 2D Full , 2001, Electrophoresis.

[4] J R Kettman,et al. Global analysis of gene expression in cells of the immune system I. Analytical limitations in obtaining sequence information on polypeptides in two‐dimensional gel spots , 2000, Electrophoresis.

[5] Kelvin H. Lee,et al. The scaled volume as an image analysis variable for detecting changes in protein expression levels by silver stain , 2001, Electrophoresis.

[6] Anders Blomberg,et al. Interlaboratory reproducibility of yeast protein patterns analyzed by immobilized pH gradient two‐dimensional gel electrophoresis , 1995, Electrophoresis.