anyone has idea to raise antibody, how much protein need? - (Aug/01/2005 )
as the topic, how much protein need for raising antibody. I think not that much. my boss wants me use FPLC to get huge amount of protein to do it. i think it is not necessary. anyone has anidea?
it depends - as always!
for immunisation of mice we use three times 10µg antigen together with adjuvants and get nice polyclonal sera (up to 1ml of serum when animal is finally sacrificed)
for immunisation of rabbits we used thrice 500µg protein....(you'll get 50-100ml serum/rabbit enough for purification of polyclonal antibodies)
for immunisation of rats (to generate spleenocytes for fusion monoclonals), we used thrice 25µg antigen....
you'll need additional antigen for testing of titers, antibody-binding assays, epitope mapping etc. though...