SDS quantitation and removal with resins

Scott Coutts scott.coutts at med.monash.edu.au
Mon Jul 7 17:35:09 EST 2003



Dr Engelbert Buxbaum wrote:
> Scott Coutts wrote:
> 
> 
>>Hi!
>>
>>Does anyone know of a way that I can assay for SDS concentration in a
>>sample containing protein? I want to know how much detergent remains 
>>after attempts at removal. I need to assay for regular (non-labelled) 
>>SDS, by the way.
>>
>>Also, I'm thinking of removing the SDS from some protein samples using a
>>resin, perhaps Dowex 1-X2 or 1-X8 or similiar. 
> 
> 
> You can assay for any detergent above cmc by measuring the fluorescence
> of DPH (1,6-diphenyl-1,3,5-hexatriene), which is non-fluorescent in
> water but highly fluorescent when dissolved in micells (358/450 nm).
> This is also a nice way to measure the cmc (Chattopadhyay & London,
> Anal. Biochem. 139 (1984) 408-412).
> 
> For the removal of detergent from biological samples you can use
> hydrophobic beads like BioBeads SM-2 (Philippot et al., BBA 734 (1983)
> 137-143), there are also various recent papers by the group of Rigaud
> which are worth reading, for example Eur. Biophys. J. 27 (1998)
> 305-319). Pierce has a similar product, packed in little disposable
> columns.
> 

Hi,

THanks for the info. I think the >CMC fluorescence method is as close as 
I'm going to get to an assay method... unfortunately, I dont think it's 
going to be usefull for me.

I agree that the hydrophobic bead method may be the best bet for 
removal. I knew about the pierce product already (if you mean the 
SDS-Out product), and had wondered what they use in that. They say it's 
a 'precipiation reagent'... but the spec sheet and instruction 'manual' 
on their website doesnt really give any information about whats in it or 
how it works. Have you used it before? You say that it's column-based; 
does it use hydrophobic beads in that or do they rely on a (at leatst 
semi-) specific precipitation of SDS?

Anyhow, thanks for the info.




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