dilution of antibody

Cheryl Smith cheryla.smith at utoronto.ca
Mon Feb 21 12:36:23 EST 2000


Hello out there,

One of the users of our facility has a concern regarding the advisability
of diluting a particular antibody.  Since these reagents are quite
expensive she doesn't want to go ahead assuming that all will be well.
Please respond to her following question directly at:
sandra.aswald at utoronto.ca

Thanks

I am planning to use the antibody:  anti-human IL-12 (p40/p70) mouse IgG FITC;
cat.# 18994A from Pharmingen.  It should be used at aprox. 0.5ug per 10
million cells, and comes as 0.1mg in a total volume of 200ul.  I was
wondering
a) use only 1ul undiluted AB per 10 million cells
b) dilute 1:10 and use 10ul

Question:
.Since I will be doing intracellular FACS with permeabilized cells, is it
O.K. to dilute the antibody?
.If I can dilute it, is there an alternative to using the Pharmingen
Perm/Wash (TM) buffer?  This is the only thing that Pharmingen suggests
diluting their anti-cytokine AB's with.

Thanks for your help
Sandra





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