IPG strip,Denaturing slab gel,Denaturing See reference: Proteomics (2006) 6:2422 See URL: http://www3.interscience.wiley.com/cgi-bin/fulltext/112511043/PDFSTART

Analytical gels were scanned on an Image Scanner (GE healthcare, Uppsala, Sweden) at 300 dpi with 12-bit gray scale levels in tagged image file format (TIFF), and images were analyzed using the ImageMaster 2D Platinum (GE healthcare, Uppsala, Sweden). All gels in the analyses were scanned with identical parameters. In order to perform differential analysis of the 2D gels with the ImageMaster 2D Platinum software, the individual spots of each gel were detected by their boundaries, and the spot volume corresponding to the protein abundance was calculated automatically. Each spot intensity volume was processed by background subtraction and total spot volume normalization; the resulting spot volume percentage was used for comparison. Manual editing of the gels was necessary especially in the higher molecular weight region. Only protein spots that were reproducibly different in all three experiments by at least a factor of two were considered to be significant and were excised from gels for analysis by MS. [Protocol text in here] [Protocol text in here]