> ---------- > From: Jose Benito > Sent: Friday, 19 May 2000 2:40 > To: Cytometry Mailing List > Subject: annexin-V stainning > > > Hi everyone: > Has anyone out there done annexin-V staining in whole blood, without > isolating PBLs?. I am trying to do it (in combination with other surfarce > markers) but I am having problems and I think that the lysing solution I´m > using is altering the results of the stainning. > Thanks > Jose Miguel > ________________________________________________________________________ > Get Your Private, Free E-mail from MSN Hotmail at http://www.hotmail.com > > -------------------------------------------------------------------------- > -------------------------------------------------- > > Hi Jose, > > Annexin staining works fine in whole blood lysis techniques if you add > some Ca + Mg ions. Likewise with your subsequent washes. You can even > permeabilise your cells following Annexin staining provided these ions are > present. When using whole blood lysis techniques you can also stain with > Annexin following other surface staining- give your cells a final wash in > your Ca/Mg buffer, then stain with Annexin, then collect your data, > although in my hands the Annexin stain works better if you stain prelysis. > The following reference may help "Optimal storage conditions for > preserving granulocyte viability as monitored by Annexin V binding in > whole blood. G.Hodge et al, J.Immunol. Meth. 225 (1999) 27-38. > > Regards, > Greg Hodge. > (hodgeg@mail.wch.sa.gov.au) > >
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