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T. A. Bogush, M. V. Tikchomirov, E. A. Dudko, M. N. Sinitsyna, R. J. Ramanauskaite, B. E. Polotsky, S. A. Tyulyandin, M. I. Davydov

Quantitative immunofluorescence estimation of Pgp expression in human solid tumors by flow cytometry


An immunofluorescence assay that enables quantitative detection of level and intensity of multidrug resistance marker Pgp expression in human solid tumors by using flow cytometry and monoclonal antibody (clone 17F9, BD Pharmingen) was developed and adapted for clinical assessment. The analysis includes the following relevant task-specific stages. Single-cell suspension can be prepared from both surgical biopsy native tumor tissues and tumor specimens fixed in 4% formaldehyde. The assay allows to result verification even after two years of tumor material storage. Cost of analysis can be decreased by increasing of period of cell incubation with antibody and reducing of the antibody concentration. It was shown also a possibility to analyze minimal tumor samples, e.g. endoscopic biopsy specimens, by decreasing concentration of cells incubated with antibodies to 1x105 cells/ml, and flow cytometer counted cell number to 1x103. Although its continuance, the developed assay does not require any changes in investigator work schedule.
Moscow University Chemistry Bulletin.
2012, Vol. 53, No. 3, P. 207

Copyright (C) Chemistry Dept., Moscow State University, 2002
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