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eluting and antigen from antibody-agarose

vasvale at yahoo.com vasvale at yahoo.com
Mon May 13 13:07:24 EST 2002


Hi
I'd appreciate if you could give me any suggestion on this technical
problem. I have to elute an antigen from an antibody agarose to do a
mass spec analysis of the immunoprecipitate. I tried  to elute the
antigen from the antibody but  didn't get complete elution. I tried both
 1M acetic acid, 50 mM ammonia and 100 mM TEA at pH 10.9.  This might
reflect the fact
that I'm immunoprecipitating with polyclonal abs and maybe some of them
have a very high affinity.  The way I did the elution is by adding 200 ul
waiting 5 min in ice, spinning and again 3 times. 
thank you if you can give me any suggestion
Valeria

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