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    Reason for protein spread throughout sucrose gradient?
    Posted by: VDEP (IP Hidden, New member, 3)
    Date: September 25, 2007 08:21PM

    I have begun to use discontinuous sucrose gradients to fractionate a 200,000g crude membrane, 1% TX100-insoluble pellet. I am examining three subtypes of neurotransmitter receptor and find that one stays at the bottom of the gradient, one is in the 200,000g soluble fraction and the third is spread throughout the gradient, from the bottom to the fractions where flotilin localizes.

    Can anyone explain the appearance of that particular receptor throughout the gradient? I doubt it is an overloading problem, since I re-ran the gradient using 5 X less protein, and saw exactly the same pattern.

     

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