[Histonet] RE: protein concentration on tissue culture supernatant

[Milne, Katy]

Hey Christina, 

I'm not sure how much the Bradford will help as there will likely be a lot of other proteins in the medium that the antibody is in (ie FBS etc) as you mention.  You wouldn't just get the Ab concentration.  Maybe for some biotinylation kits this isn't a problem but I'm not sure what you're using.

Do you have a kit that you can straight up biotinylate the Ab while it's still in the medium or do you need to purify it?  Does it need to be biotinylated or can you come in with a biotinylated secondary (or are you dealing with mouse-on-mouse problems)?

Katy

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Date: Thu, 11 Oct 2012 15:05:39 -0400
From: "Thurby, Christina" <christina.thurby <@t> bms.com>
Subject: [Histonet] protein concentration on tissue culture
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Please don't blast me if this is a really dumb question.  I have a tissue culture supernatant antibody.  I need to know the protein concentration - called the vendor, it is unknown.  Does anyone know if I can just run a Bradford to get the protein concentration - or will it be irrelevant do to other proteins present in the antibody?  This is an IVD antibody, so I expect it to be specific for the target antigen.  This is an unconjugated antibody and I want to biotinylate it, but I need a starting protein concentration.  Anyone out there with experience doing this with a tissue culture supernatant antibody?

Christina Thurby
Bristol Myers Squibb
812-307-2093