Mouse serum incubation prior to staining

arjmand arjmand at farabi.tums.ac.ir
Mon Sep 18 23:43:07 EDT 2006


Dear Liza

For blocking the nonspecific binding sites, one has to use large volum amount of mouse
serum if purified IgG is not avalable. In this case the cellular concentration as well as
protein concentration will be changed, so for adjusting them again you'd better wash them
bafore the immunofleuorescent staining.

yours,

Alireza Ardjmand

Hematology, Oncology 
and B.M.T research Cent. 
Shariati Hospital 
Kargar Ave. 14114 
Tehran-Iran

alternative email: 
ardjmar72 at yahoo.com

www.tums.ac.ir

On Sat, 16 Sep 2006 18:40:49 -0400, mcguirel wrote 
> Hello Flowers, 
> 
> Quite some time ago I was introduced to incubating the stem cells 
> with Mouse serum prior to staining in order to minimize NSB.  This 
> incubation was always followed by 2 washes prior to the actual 
> staining of the cells with the antibodies in order to rinse out any 
> excess serum. 
> 
> Last week I was informed by a fellow flower that this is no longer 
> necessary. The protocol would be to incubated the cells with mouse 
> serum for 10 minutes and then add the antibodies directly in the 
> mixture without any washes (then continue with lyse/wash/fix). 
> 
> What are your thoughts. Can anyone corroborate this? 
> 
> Thank you very much for your help. 
> 
> Liza McGuire 
> 
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