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Calculating amount of Beads for purification of Ig


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#1 Sharmistha Das

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Posted 28 February 2013 - 08:55 AM

Hi, I am using Milipore's magnetic protein G beads to purify monoconal antibodies. Can anyone help me with the calculation as to how much beads to use? The binding capacity for these beads are 2.5-3.5mgs/ml of suspension and I am using supernatant from hybridoma cells.
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#2 Curtis

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Posted 28 February 2013 - 10:50 AM

I'm not sure about the binding capacity but I used 10 to 30 ul for my work. Even 10 ul purified a lot.

#3 Sharmistha Das

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Posted 28 February 2013 - 06:19 PM

Hi Curtis, Thanks so much. Could you tell me how much your starting material was ie how much supernatant from hybridoma cells you used for your work? And did you concentrate your supernatant? Thanks

#4 Curtis

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Posted 28 February 2013 - 09:07 PM

I did not concentrate. I added 100-1000 ul supernatant. It will bind, don't worry.




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