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    SDS-resistant protein aggregation
    Posted by: Mari (IP Hidden, New member, 2)
    Date: March 21, 2006 11:23AM

    If an hight concentration of a protein give rise to an aggregation due to a non covalent bond SDS-resistant, what could I add to the sample preparation (for an SDS-PAGE analysis) in order to avoid it?
    I mean: if this bond is SDS-resistant, what else can break it?


    Thanks in advantage!

     

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    Re: SDS-resistant protein aggregation
    Posted by: supernov (IP Hidden, New member, 3)
    Date: April 20, 2006 08:57AM

    Is the protein a result of an induction? It could be that it's in inclusion-bodies which are very resistent to this. I believe sonication could help.

     

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    Re: SDS-resistant protein aggregation
    Posted by: femmeauburn (IP Hidden, Advanced member, 115)
    Date: July 19, 2006 03:22PM

    Do you add DTT or mercaptoethanol to your loading dye? You can also boil the samples to aid in denaturation, which should help break the bonds.

     

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    Re: SDS-resistant protein aggregation
    Posted by: coco (IP Hidden, New member, 1)
    Date: July 20, 2006 06:38AM

    If your sample is inclusion body, homogenize precipitation with 8M urea, befor you treat the protein with sample buffer (contained SDS) . Most proteins can be dissolved.

     

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