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sucrose gradient for protein fractionation?


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#1 minylim

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Posted 12 July 2010 - 09:41 AM

Hello,

I would like to have fractions of my whole cell extract, I'm trying to isolate nucleoplasmic and cytoplasmic fraction.
But my PI insists that I can use sucrose gradient for fractionation of my cell lysates.
I thought it is just for organelle fractionation and cannot be applied to my sample, which is whole cell lysate sup after spin.
Gradients kit contains from 8.5 to 60 percent sucrose.
Someone please help whether I can use the method. Thank you in advance!!

#2 Inmost sun

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Posted 13 July 2010 - 01:22 AM

View Postminylim, on Jul 12 2010, 05:41 PM, said:

Hello,

I would like to have fractions of my whole cell extract, I'm trying to isolate nucleoplasmic and cytoplasmic fraction.
But my PI insists that I can use sucrose gradient for fractionation of my cell lysates.
I thought it is just for organelle fractionation and cannot be applied to my sample, which is whole cell lysate sup after spin.
Gradients kit contains from 8.5 to 60 percent sucrose.
Someone please help whether I can use the method. Thank you in advance!!

of course, you can fractionize proteins by sucrose density ultracentrifugation; it works best for pre-purified fractions and macromolecular proteins f.i. oligomeric proteins or large fibrillar proteins




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