Detection of Cy5

system@flocyt.int-med.uiowa.edu
Tue, 07 Nov 1995 10:12:37 EST

The folks from Australia had two questions about using Cy5 =>

a. Ease and stability of conjugation
b. Intensity of fluorescence

We use Cy5 conjugated antibodies on a dailey basis excited by a dye
laser tuned to 600nm and emitting 400mW of power. The intensity of
fluorescence is excellent and may possibly be better than APC. We
have never done a direct comparison. Cy5 is the third color of choice
here at the Univ. of Iowa because of its ease of conjugation and good
fluorescence characteristics. We routinely run fluorochrome combinations
of FITC, PE, Cy5, and PI for live/dead discrimination and four color
experiments using FITC, PE, Texas Red, and Cy5. The people who do
Cy5 conjugations here tell me that it is as easy as conjugating FITC
and I have not heard any complaints about stability.

In our hands Cy3 does not work as well as PE. The fluorescence
intensity is much lower.

Hope that helps

Justin Fishbaugh
University of Iowa
Flow Cytometry Facility

internet: justin-fishbaugh@uiowa.edu


Home Page Table of Contents Sponsors Web Sites
CD ROM Vol 2 was produced by 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:(317) 494-0757; FAX (317) 494-0517; Web http://www.cyto.purdue.edu EMAIL robinson@flowcyt.cyto.purdue.edu