[Cell-biology] Dissolving lysophosphatidic acid

Anita Umesh via cellbiol%40net.bio.net (by aumesh from stanford.edu)
Tue Oct 14 19:27:51 EST 2008


Hi, I need some help dissolving lysophosphatidic acid so that it can 
be applied to cells.  It seems to vary each time it is dissolved 
(final solution contains calcium (2mM) and magnesium (1mM).  I am 
first dissolving it in a solution that contains fatty acid free bsa 
(1mM LPA in 1mM BSA containing PBS).  I am sonicating this solution, 
as well as the calcium and magnesium containing final buffer.  but it 
seems that the response (Calcium imaging using fura-2) that I am 
getting from the cells is variable - also seem to have to use pretty 
high concentrations >10uM in order to get a robust response.  I 
notice that at higher concentrations of LPA, that my chamber 
containing the cells and the solution has precipitates in it by the 
time I am done imaging.

Since people have used this to get calcium transients from cells, 
there must be a trick that I do not know about.

Please help!
Thank you,Anita
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