Please note that this approach (blocking with BSA-Cy5) will only work if
you are not using a second laser with APC as a stain. If you use APC the
compensation would become impossible (i.e. the background staining of the
bound BSA-Cy5 would wreak havoc on the APC channel)..
Alan Stall
BD-PharMingen
===================
Gina_Graziani-Bowering@hc-sc.gc.ca on 04/17/2001 10:54:36 AM
To: cyto-inbox
cc: (bcc: Alan Stall/SDCA/BDX)
Subject: Re: PE-Cy5 Backgrounds
Hi Doug,
To avoid the problem of PE-Cy5 conjugated antibodies binding to Cy5
receptors on the surface of monocytes, we used to preincubate the cells
with
BSA-Cy5 reagent prepared by conjugating BSA (Sigma) to Fluorolink Cy5
Reactive
Dye
(Amersham Life Science, Inc., Pittsburgh, Pa.) as per the manufacturer's
instructions. This process is analagous to blocking cells with gamimmune
prior
to staining. Hope this helps,
Gina
_______________________
Gina Graziani-Bowering, PhD
Post-Doctoral Fellow
Health Canada
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