How to remove RNase from immunoprecipitation?

We are trying to do a "simple" immunoprecipitation in which
the unbound fraction will be assayed for RNA processing activity. We find that
protein A-agarose beads have little non-specific RNase activity. After adding
serum (immune or non-immune), we wash the beads up to 10 times until the wash
supernatant has no detectable RNase.


However, when we do the actual IP by incubating these beads with the experimental enzyme fraction, the unbound supernatant has much more non-specific RNase activity ... ...

Last update 03-Dec-2003, Rating Good of 0 votes.