Rapid production of quasi-stable antibody-phycoerythrin conjugates for use in flow cytometry

✍ Scribed by George L. Dale

• Publisher: John Wiley and Sons
• Year: 1998
• Tongue: English
• Weight: 75 KB
• Volume: 33
• Category: Article
• ISSN: 0196-4763
• DOI: 10.1002/(sici)1097-0320(19981201)33:4<482::aid-cyto13>3.0.co;2-j

✦ Synopsis

A simple system is described for producing quasistable 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 streptavidinfluorochromes can be used simultaneously. Cytometry 33:482-486, 1998.