re: Annexin V

From: C.Jacobs@nefro.azn.nl
Date: Thu Mar 04 1999 - 03:55:47 EST


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Hi, we have problem in labeling cells with Annexin V. In our protocol the
cells are resuspended in an incubation buffer which contains 1.5mM CaCl2,
then we add 5 ul of annexin V and we leave on ice for 15'. 
With this protocol we did not observed any positive cell even in the
positive control. Could  anybody give us suggestions?
thank you
Silvana   


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Hi Silvana,

Increase the Ca concentratie up to 2.5 mM.
Normally the Ca conc. is between 1.8 - 2.8 mM.
There is a possibility that you get a precipitate of Ca-X
[phosphate]. Than take another buffer/medium.


Cor



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