polyclonal antibodies using inclusion bodies - how to prepare sample (Feb/25/2008 )
i'm new in production of polyclonal antibodies field..currrently i'm trying to generate polyclonal antibodies using inclusion bodies from my recombinant protein.. i'm still searching for the best way to prepare the inclusion bodies samples to inject into the rabbit..is there anyone out there can help me?plz...
the inclusion bodies are his-tagged recombinant protein produced in e.coli..do i have to solubilize the pellet using the solubilization buffer (8M urea, 100mM NaH2PO4, 10mM Tris). but i'm not sure whether urea will give effect to the rabbit?
thanks!!!really appreciate your help!!
I presume you are actually interested in purified protein, rather than the inclusion body, per se. If so, the solubilization buffer you propose sounds fine. Once you have purified the protein on a nickel column, I'm guessing, you will need to refold it, otherwise the polyclonals you make will be against denatured and refolded protein (unless that is you intention...).
For information on refolding go to the Monash University Refold database (http://refold.med.monash.edu.au/).