SDS quantitation and removal with resins
Dr Engelbert Buxbaum
engelbert_buxbaum at hotmail.com
Mon Jul 7 10:07:13 EST 2003
Scott Coutts wrote:
> 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
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