staining of cord blood CD34+ cells

Yashpal Agrawal yagrawal at
Thu Nov 21 08:52:12 EST 1996

Dear flowusers,
		Problem:I have been having problems with CD34 cell
staining ofwhole Cord Blood(CB). Specifically the Isotype control (IgG1-PE from
 B.D) gives non-specific staining (0.2-0.6 % of mononuclear cells). This
small amount of non-specific staining causes difficulty in interpretation
when the CB stained with the specific HPCA-2-PE (also from BD) gives a
positivity in  0.5 % of cells and subtracting the background gives 0%

Do others have the same problem, any suggestions/advice is welcome ?.

Methods used:We collect our CB in blood bags (CPDA) and the sample is
stored at 4 C until  analyzed which is 1-4 hrs. Staining is according to
standard methods. 50 ul of CB + 20 ul antibody is incubated for 15 min in
cold and in dark. After this 2 ml FACSlysis solution (BD) is added,
incubated for 10 min and centrifuged for 5 min at 400 g. The cells are
then washed twice in PBS and analyzed immediately. 

Thanking you all,
Yash Pal Agrawal
Kuopio, Finland
yagrawal at

More information about the Cytometry mailing list