Re:Coupling of proteins

tarnok@medizin.uni-leipzig.de
Thu, 18 Jul 1996 09:11:45 +0000

Dear Matthias,

we have used succesfully microbeads from Covalent Technology
Corporation (Covaspheres). They provide beads for antibody and
protein or avidin and lectin coupling and work very nicely. The
peptides are coupled chemically to the beads. The beads are 0.5um in
diameter and have quite good CVs of around 2-5%. They also provide
larger and smaller beads but the 0.5um worked very well. See for
details: J. Cell Biol. 130(2):473-484 (1995), Neuron 11:1113-1122
(1993), Neuron 10(4): 711-727 (1993). Recently we turned to Dioclean
TM from Duke Scientific (Palo Alto CA) (also 0.5uM beads). The
coupling of peptides to these particles is very simple as it works by
hydrophobic interaction. CV etc are similar to those of the
covaspheres.

If you want to measure protein interaction on beads by flow (as we
did) less than 0.5um is bad for triggering by FCS and at least in my
hands the resolution of the aggregates decreases. Blocking of free
sites was done with BSA.

I hope this helps

Attila
****************************************************
Dr. Attila Tarnok phone:**49-3418652430
Clinics for Pediatric Cardiology fax: **49-3418651405
Heart Center Leipzig GmbH
University Hospital, Russenstr.19
D-04289 Leipzig, Germany
Internet:tarnok@server3.medizin.uni-leipzig.de
****************************************************


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