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The best flow cytometry substrate for the detection of lacZ activity
is a lipophillic FDG derivatves with 2, 4, 8 and 16 carbon acyls
known as ImaGene (Molecular Probes). However, this substrate has a
problem for the quantification of lacZ activity. We have used it to
assay b-gal activity in insect cells infected with baculovirus. A concentration
equilibrium between cell and supernatant of the hydrolysed substrate,
irrespective of enzyme activity, meant that lacZ+/lacZ- resolution
was impossible. Quantitative measurements proved problematic because the
enzyme activity showed no sign of saturation by the substrate.
Good luck
Mohamed Al-Rubeai, Ph.D.
School of Chemical Engineering
University of Birmingham
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web