preferred method for purifying antibodies - (Jan/10/2008 )

What is your preferred method for purifying polyclonal antibodies?

Currently what our lab does is this: run a large amount of purified antigen on an SDS-PAGE gel and blot to nitrocellulose, stain with ponceaus, cut out a strip containing the antigen , probe with about 300 ul of antibody serum, wash several times and elute with about 300 ul of an appropriate elution buffer.

This procedure works ok, but it's time consuming and not always efficient. Plus my antibody is not very stable using this method. Only lasts about a month at 4C.

I'm wondering if anybody has a tried and true method that they swear by.

Thanks!

Ammonium sulphate precipitation - because it's cheap and this purity suits well my purpose.
My friend is using protein A or G - more expensive but very good.

If you really need antigen-antibody affinity purified antibodies, you may consider making Sepharose column with conjugated antigen, instead of method you use now.

Ammonium sulphate precipitation - because it's cheap and this purity suits well my purpose.
My friend is using protein A or G - more expensive but very good.

If you really need antigen-antibody affinity purified antibodies, you may consider making Sepharose column with conjugated antigen, instead of method you use now.