Background: Dissociable Antibody MicroArray (DAMA) staining technology is a new approach for global analysis of protein expression and subcellular localization profiles in fixed cells. We have developed the technology and demonstrated its application on identifying differentially expressed proteins in breast cancer cells. We have determined the subcellular localization profiles of 360 proteins in breast cells and identified a protein with unique subcellular localization in breast cancer cells, by combining data from four 96-antibody format arrays. In this report, we have further developed the technology to determine the subcellular localization profiles of 400 proteins simultaneously from single chip, in a 400-antibody format.
Method: We have determined the subcellular localization profiles of 400 arrayed antibodies in two normal prostate cell lines (PWR-1E and PZ-HPV-7), and three cancer cell lines (Du145, LNCap and VCap). The subcellular localization profiles were compared and analyzed by Chipview, the program for image database management and analysis.
Results: A protein, GRK2 was identified to have unique localization in some of the cancer cells: localized in the membrane of two cancer cell lines, and in the cytosol of normal cell lines and one cancer cell line. The identified subcellular localization difference was confirmed by individual immunostaining.
Conclusion: DAMA staining technology could be a powerful method for global subcellular localization profiling in high throughput format.