RE: annexin-V stainning

From: Hodge, Greg (HAEM) (hodgeg@mail.wch.sa.gov.au)
Date: Thu May 18 2000 - 16:59:48 EST


> ----------
> 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
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> --------------------------------------------------------------------------
> --------------------------------------------------
>
> 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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