I might have overlooked something, but I could not find any information on what the maximum concentration of sodium deoxycholate in my buffer can be for StrepTagII-Streptavidin purification.
In my sample, I have 1% DOC and I cannot dilute it out because this will facilitate protein aggregation.
Does anyone have experience with this?
I might be able to replace the DOC with SDS or NP40 if DOC has to be lowered.
Thank you!
Hello,
I don't know much about DOC or NP40, but you may have a look at this:
http://www.iba-lifesciences.com/tl_files/uploads/bilder/produkte/streptag/Downloads/Reagents%20compatible%20with%20Strep-tag.pdf
I think that if you want to use DOC, you may try and see what happens, or you can contact the technical support of IBA, they usually answer quite fast!
Hi Simone,
thank you for your answer. I already had this list, but I might try contacting the support. And also just give it a shot with the 1% DOC I have as well!
Hi Chiara, is there a reason why you need to use streptavidin rather than StrepTactin? Can you also increase the salt concentration to keep your protein soluble (or add proline) rather than use DOC?
Anyway, the IBA list includes a number of ionic detergents, including SDS. Perhaps begin by determining the relative molecular weight of SDS to that of DOC, as well as the charge relative to that of DOC, to estimate how high a DOC concentration (% w/v) you might be able to use.
Nonidet P40 and NP-40 (nonylphenoxypolyethoxyethanol), unlike DOC, are both non-ionic detergents so I don't think they will work well as a substitute if you are depending on DOC.
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