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background problems with Abs

Susan Mango smango at macc.wisc.edu
Sat Apr 22 19:00:02 EST 1995


i've used worm acetone powders and it's helped (absorbed both the
primary and the secondary). generally if there is little binding of the
primary i've had more problems with the secondary having background.
but this tends to be rare. blocking for a longer time could make a
difference and washing the primary well.... we buy our secondaries from
jackson (donkey anti--, not goat) and i tend to use rhodamine
conjugated Abs because they don't bleach as quickly. knocking down the
concentration of your antibodies might also help (i use the secondaries
at 1:150-200). and sometimes secondaries can go bad (although it's
rare). let's see....can't think of anything else off-hand -- susan



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