 |
 |
 |
 |
We (the royal We) made several attempts to work with sodium green
with limited success. The limit was that we could get it into the
cells (Jurkat, a line loosely referred to as a T-cell) but we saw
no change in fluorescence in the presence of either veratridine or
gramicidin. We tried different concentrations of S.G. (1-5 uM),
different temperatures, different times of loading and stimulation,
etc. The only thing we noticed was that cells had a tendency to die
the longer they sat. It was at that point that we gave up and thought
about trying SBFI in a different system. After a very informative
conversation with a colleague of mine I learned that SBFI and PBFI have
only about twice the affinity for their own ion (Na for SBFI, K for PBFI)
as they do for the opposing ion (K for SBFI, Na for PBFI) not an ideal
situation if you are trying to measure changes without first clamping one
or the other. I wonder how Sodium Green compares in affinity. Our interest
in looking at sodium fluxes waned (although it has not disappeared) at
this point so we went off to pursue other interests without ever testing
the SBFI or PBFI. As I say our interest has not disappeared so if anyone
has any better luck or good suggestions please post them.
Thanks,
Keith
**********************************************************************
*
Keith A. Kelley kelley@mrc.com * A wink is as good
Miles Research Center Ph 203-937-2872 * as a nod to a
400 Morgan Lane Fax 203-937-6923 * blind horse.
West Haven CT 06516 *
*
**********************************************************************
|
|
|
|