Hi,

I am working with a user on neurons culture (embryonic) in order to
characterize a list of antibodies for differentiated and undifferentiated
markers. Here the problem: either transcription factor (nuclear) and
membrane antigens show staining in the nuclei and in the membrane. it
seems the antibody sticks on the membrane or enter partially in the
nuclei.

The protocol is 4%PFA 10min RT, triton 0.3% 10min, bl0cking in Normal goat
serum,and washes in TBS-tween0.1%.

Any suggestions?

Thanks in advance

Valeria


Berno Valeria, PhD

EMBL- Mouse Biology Unit
Campus A.  Buzzati-Traverso
Via Ramarini,  32
00015, Monterotondo Scalo  (RM)
Italy

Tel:  +39 06 90091287
Fax: +39 06 90091406
email: valeria.berno@embl.it
www.embl.it


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