We use the kit from Calbiochem but ran out of binding buffer somehow. We have never prepared one before.
Here is one for 10x : 0.1 M HEPES/NaOH, pH 7.4; 1.4 M NaCl; 25 mM CaCl 2.
1. THIS IS 10X. Dilute to 1x before use. I generally just make 1x and filter; you can store it for quite some time.
2. Calcium is very important for binding. Add it from a stock solution or it may precipitate if added as powder.
Good luck !
Thank you Prem I let you know! But what is the need for such components?
The high CaCl2 in the common AnnexinV binding buffers can be toxic to some cell types.
Here is a simple alternative buffer:
2.5mM CaCl2, 140 mM NaCl, 4 mM KCl, 0.75 mM MgCl2 and 10 mM HEPES in DDW
For more information follow:
http://www.cyto.purdue.edu/cdroms/cyto10a/educationandresearch/annexin.html
As my understanding, it is critical to have calcium/magnesium in the buffer for functioning of the Annexin V and to keep pH correct. Is it possible to use HBSS or PBS supplemented with calcium/magnesium for the same purpose? pH is preserved and Ca/Mg supplemented. SHall it work?
Dmitro: I used Pred Submariaman below and it worked like a charm in Vero and Hela cells.
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