Re: Annexin V

From: japhilip (Jan.Philippe@rug.ac.be)
Date: Fri Apr 30 1999 - 04:44:39 EST


> --On Fri, Apr 23, 1999 9:45 AM +0000 Rita Cavaleiro
> <ritacavaleiro@hotmail.com> wrote:
> 

> >  we obtain, unexpectedly, a lot of positive cells for annexin in 
> > unstimulated human PBMCs in culture for 24 hours. I am not sure if 
> > this high level represents unspecific labelling because there is no 
> > wash after the adition of annexin. None of the protocols  use this 
> > wash. Could anyone tell me if there is any problem in doing this wash? 
............ What is the best positive control to induce apotposis in 
> > human PBMCs and in what concentrations ?. I tried Dexamethasone but it 
> > did not work.
> >
a 'background' after 24h in vitro culture of about 7% is normally 
found (if incubated in 'optimal' conditions, e.g. with FCS 5%).
A possibility for positive control is gamma-irradiation. More data 
can be found in Cytometry 1997; 29: 242-9.
Jan Philippé
e-mail: jan.philippe@rug.ac.be 



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