4 C labeling might accomplish a couple of things. With live cells, they are in a constant state of change, as a metabolic organism is prone to be. Keeping them at 4 C will maintain their phenotype and most likely keep more of the cells viable (death requires energy). 4 C labeling should also decrease non-specific binding of your Ab. I know some people who goes as far as doing all labeling in a cold room, so as to avoid taking the antibodies from 4 C to 25 C and back again. Good luck. Keith Bahjat kbahjat@ufl.edu On 3/12/01 4:18 PM, "Simon Monard" <smonard@trudeauinstitute.org> wrote: > > Hi > > People always used to incubate on ice for 30-60 minutes to try to prevent > capping, also > to create a convenient time to have lunch or extended coffee and smoking > breaks. Nowdays > you can add a tad of Sodium azide to prevent capping. > > Simon > > Simon Monard > FACS Lab Manager > Trudeau Institute > Saranac Lake > NY12983 > > Ph 518 891 3080 X352 > > >>>> "Heather Medbury" <Heather_Medbury@wsahs.nsw.gov.au> - 3/11/2001 5:50 PM >>>> >>> > > Hi FLOWers, > > About 5 years ago when I was analysing lymphocytes, we used to have the cell > preparations > at 4 C for labelling with the antibody. > The work I do now on whole blood, the cells are labelled at room temperature. > > Why do people label at 4 C when it works perfectly well at room temp? > > Heather > > > > Heather Medbury (PhD) > Department of Surgery > Westmead Hospital > Westmead 2145 > 61-2 9845 7677 > -------------------------------------------------------------- > All experiments are preliminary > > ------------------------o0)(0o-------------------------------------- > > "In his heart a man plans his course > but the Lord determines his steps" > > --------------------------------------------------------------- > >
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