polyclonal for IP - how many? (Oct/18/2006 )

hi
I´m going to do an IP using a kit from Pierce (ref.45213) and a polyclonal antibody. I can´t know the concentration of my antibody since it is whole serum from rabbit. Any idea of how many I should use to preparate the complex with the antigen? In what proportion in relation to the sample? Any idea?

And another thing: since my sample is human serum, should I remove albumin to enrich it in the lower abundance proteins?

thanks

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normally, BSA should not disturb IP beside if it unspecifically bind Ab or your protein of interest but this you will see; more problems should arise from immunoglobulins of serum; see my other answer to your similar question

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thanks a lot kosmodrom

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there is not many options : I would try several quantities from 5 to 50 µL (depending of the volume of antibodies you have), with a known positive sample (or a sample to which you add a recombinant protein), and try to find the lowest volume that is working.
good luck

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THANKS missele, you all are so useful and I´m beginner who sometimes feels so lost.

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