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SDS-resistant protein aggregation
Posted by: Mari (IP Hidden, New member, 2)
Date: March 21, 2006 12:23PM
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!
Re: SDS-resistant protein aggregation
Posted by: supernov (IP Hidden, New member, 3)
Date: April 20, 2006 09:57AM
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.
Re: SDS-resistant protein aggregation
Posted by: femmeauburn (IP Hidden, Advanced member, 115)
Date: July 19, 2006 04:22PM
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.
Re: SDS-resistant protein aggregation
Posted by: coco (IP Hidden, New member, 1)
Date: July 20, 2006 07:38AM
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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