Inespecific binding

Roxana Schillaci rschilla at
Fri Aug 14 08:59:19 EST 1998

Hi everybody:
I have a problem with indirect immunofluoresce labeling plus direct one.
When I labed Jurkat cells with an indirect immunofluorescence (first
Ab is a  mouse m Ab (IgG1) and 2nd Ab is goat anti-mouse IgG-FITC)
and then with CD25 R-PE there is a non specific binding of the last Ab
(I have 75% possitive!!) (of course I wash in each step...)
When I label the cells only with CD25 I have 3-5% possive cells, which is OK with the literature.
What is going on?
How can I ride of the non specific binding?
Someone have simmilar problems?
Thank you for helping me.

Roxana Schillaci, PhD
Instituto de Biologia y Medicina Experimental
Vuelta de Obligado 2490
(1428) Buenos Aires

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