Phosflow antibodies

Dear all.

I am planning to purchase some BD phosflow antibodies, as I didn't have
much luck with establishing phospho-specific westernblots so far in
our laboratory and am hoping to circumvent most of the problems (long
sample handling with the risk of de-phosphorylation, potential
contaminanting cells that do not belong to my target population) by
going back to the single-cell level and flow-cytometry, which is done
rountinely in our lab.

I would be grateful if anyone here could share their experiences with
phosflow antibodies with me (specifically on human monocyte-derived
DCs, that are very adherent, so I also face the problem of a long (ca.
30 min) detaching procedure following treatment - does anyone here
know if I could fix the cells prior to detaching to preserve
phosphorylation? Also, I am wondering whether the shift I will see
following treatment is in your experience really as visible and clear
as in the company's spec sheets - or very har do detect at all. I am
mainly interested in members of the MAP kinase family.

Thanks,
 Anja			       


_______________________________________________
Anja Scholzen
PhD Student

Burnet Institute at Austin
Studley Road, Heidelberg 3084
VIC, Australia
Tel.:0061 3 9287 0673
Fax: 0061 3 9287 0600