BSA following fixation and permeabilization??

From: mschabat@mail.mdanderson.org
Date: Tue May 15 2001 - 17:39:25 EST


Hello--I am an extreme novice in the field of flow, and I have a question about
an existing protocol that I am attempting to replicate.  I am trying to label a
nuclear protein (PARP).  The authors of the protocol resuspended the fixed and
permeabilized cell pellets with 1% BSA in PBS followed by incubation with the
specific FITC-conjugated Ab.  Following the incubation, the cells were pelleted
again and washed with 1% BSA-PBS to remove excess stain.  Here's my question;
what is the purpose of the BSA in PBS prior to and following incubation of the
Ab?  Also, there are a vast variety of BSA available; what would be the best in
this application?

Any information would be appreciated, Matt.

============================
Matthew B. Schabath
Predoctoral Fellow
Department of Epidemiology
M.D. Anderson Cancer Center
lab:      713-792-5760
office:   713-745-1078
fax:      713-792-0807



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