In response to Tim Wakefield, who wrote:
I am looking for some advice on lectin protocols.  I am currently
binding a
FITC-conjugated lectin to the symbiosome membrane surrounding an
endosymbiotic algal cell that has been extracted from the anemone
host
cell.  I have had good success in binding the lectins to the
membrane,
but have had NO success in inhibiting the binding through the use
of a
competing sugar.  According to published literature, the addition
of a
competing sugar to the lectin solution prior to the addition of
the
algal cells should prohibit binding to the membrane.  Well, it
just isn't
happening.  I have tried varying the concentration of lectin and
sugar
and this does not seem to have an effect.  The lectin that I am
currently
working with is succinylated-Concanavalin A and it's competing
sugar,
mannose.
 
A very potent inhibitor of Con-A (not succinylated) is alpha
manno-pyranoside and worked just fine at a concentration of 0.1M
sugar to 10-50 g/ml Con A when incubatedtogether for 30 min
before application. Sometimes incubation time is the problem.
Con A has been known to bind by ionic mechanisms also and then
the sugar wouldn't work - but I don't know if that would happen
with s-Con A. - lots of luck.
 
Libby Gretz
Univ. of Virginia
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