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IP for ceruloplasmin

IP for ceruloplasmin - Immunoprecipitation Forum

IP for ceruloplasmin - IP or Immunoprecipitation Forum.


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Old 08-17-2009, 05:19 PM
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Question IP for ceruloplasmin



Hello.

I want to deplete 15 mL of human and sheep plasma for ceruloplasmin.
I have been looking up methods but I'm confused about how do go about it.
Most people seem to follow this protocol: incubate plasma with an antibody raised against ceruloplasmin. Add IgA and/or IgG conjugated agarose beads. Incubate and pellet the beads.
But for 15 mL of plasma I would need a lot of expensive antibody.
Is there a protocol and/or product anyone can recommend? A bead system, or maybe an affinity column I can load with a specific antibody, another method? Or recommend where I should be looking for these answers?

Thank you for any suggestions.

Last edited by kvrancken; 08-21-2009 at 03:57 PM.
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Old 02-28-2013, 01:46 PM
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Default Re: IP for ceruloplasmin

Preparation of ceruloplasmin-depleted plasma
Ceruloplasmin is immunoprecipitated from human plasma by treatment with rabbit anti-human ceruloplasmin antiserum in 0.05 M TES buffer at pH 7.3 and incubating for 2 h at room temperature, followed by incubation with protein G-agarose in 0.05 M TES buffer at pH 7.3 for 2 h and centrifugation. The ceruloplasmin-deficient supernatant is concentrated by ultrafiltration using Centricon-10 filters (Amicon, Beverly, MA) and stored at -20 C. Complete removal of the ceruloplasmin is verified by SDS-PAGE on a 4-12% polyacrylamide gel followed by western blotting with rabbit anti-human ceruloplasmin antiserum.
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