Abstract
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.
Highlights
Prostate carcinoma is the most common form of cancer in men and the second leading cause of death in the United States [1,2]
Dissociable Antibody MicroArray (DAMA) staining technology could be a powerful method for global subcellular localization profiling in high throughput format
Efforts to improve the early detection of cancer by use of prostate-specific antigen (PSA) and transrectal ultrasound has result in increasing numbers of prostate biopsies submitted for examination to exclude early prostate cancer [3,4]
Summary
Prostate carcinoma is the most common form of cancer in men and the second leading cause of death in the United States [1,2]. We have developed a novel platform called Dissociable Antibody MicroArray (DAMA) staining technology [18,19,20] that combines the power of immuno-histochemical staining with the high throughput characteristics of protein microarrays. This technology provides a new approach in the global analysis of protein expression and subcellular localization profiles in fixed cells and tissues. 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 further developed the technology to determine the subcellular localization profiles of 400 proteins simultaneously from single chip, in a 400-antibody format
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