C. Muller et al., RAPID QUANTITATIVE-ANALYSIS OF PROTEIN-TYROSINE RESIDUE PHOSPHORYLATION IN DEFINED CELL-POPULATIONS IN WHOLE-BLOOD AND BONE-MARROW ASPIRATES, British Journal of Haematology, 94(3), 1996, pp. 461-463
We developed a rapid method for quantification of tyrosine phosphoryla
tion in immunophenotypically defined cell populations in specimens of
whole blood and unprocessed bone marrow. Samples were formaldehyde-fix
ed and cells were permeabilized, Phosphotyrosine residues and surface
antigens were simultaneously stained by monoclonal antibodies and visu
alized by flow cytometry. The accuracy of the method was confirmed by
demonstration of an increase of phosphotyrosine levels in pp60(v-src)
transformed fibroblasts. In blood of healthy donors, monocytes and gra
nulocytes showed higher levels of phosphotyrosine than lymphocytes. CD
34(+) peripheral blood stem cells showed slightly increased tyrosine p
hosphorylation compared to autologous lymphocytes. Significantly eleva
ted levels of phosphotyrosine were demonstrated in leukaemic blasts co
mpared to lymphocytes (P = 0.01).