A comparative study of different dyes for the detection of proteomes derived from Escherichia coli and MDCK cells: sensitivity and selectivity.
We compared sensitivity and selectivity of five dyes for detection of 2D PAGE-resolved proteins derived from Escherichia coli and MDCK cells. The sensitivity of these dyes was in the following order: SYPRO Ruby>Deep Purple>CBB-G250>CBB-R250>Colloidal Gold. Also, we report herein for the first time the application of Colloidal Gold (which is commonly used for staining proteins on blotted membranes) for in-gel staining of proteins. For E. coli, most of the dyes preferably detected proteins with pI range of 4.0-6.9, whereas Deep Purple preferably detected proteins with less acidic range (pI 5.0-7.9). For MDCK cells, while other dyes preferably stained proteins at pI range of 5.0-7.9, Colloidal Gold preferably stained more basic proteins (pI 7.0-9.9). This preferential staining property of Colloidal Gold to basic proteins was confirmed in SDS-PAGE-separated lysozyme (pI 9.4), compared to calmodulin (pI 4.0) and albumin (pI 6.0). These data provide useful information to select appropriate dyes for gel-based proteomic analysis of individual samples.
Chiangjong
Wararat
W
Faculty of Medicine, Medical Proteomics Unit, Office for Research and Development, Siriraj Hospital, Mahidol University, Bangkok, Thailand.
Thongboonkerd
Visith
V