Rapid production of quasi-stable antibody-phycoerythrin conjugates for use in flow cytometry.
A simple system is described for producing quasi-stable complexes between biotinylated antibodies and fluorochrome-labeled streptavidin. Optimal complexes were produced by mixing biotinylated antibody and phycoerythrin-streptavidin together at a molar ratio of 1:1 for 2 min, followed by the addition of soluble biotin to halt oligomerization. These complexes, which are stable for at least 4 h, exhibit fluorescence with cell binding which approximates that observed with standard two-step labeling techniques. Equally important, multiple biotinylated antibodies complexed with different streptavidin-fluorochromes can be used simultaneously.
Dale
G L
GL
Department of Medicine, University of Oklahoma Health Sciences Center, Oklahoma City 73190, USA. george-dale@ouhsc.edu
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D004285:Dogs; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D019012:Integrin beta1 / Q000276:immunology*; D010799:Phycoerythrin; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000276:immunology*; D019038:Platelet Glycoprotein GPIb-IX Complex / Q000276:immunology*; D013997:Time Factors
D000911:Antibodies, Monoclonal; D019012:Integrin beta1; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D019038:Platelet Glycoprotein GPIb-IX Complex; D010799:Phycoerythrin