Hello, I am currently working on a project related to study of the intact protein by Top Down Proteomics. Proteins are fractionated by Gelfree 8100 Fractionation Station and after fractional separation protein contain lots of SDS in solution. I have tried FASP to remove SDS but did not get much success. Acetone/Methanol ppt is also not a suitable proticol since protein quantity is very small (few ug). Is there any other way to remove SDS which can reduce the loss of protein?
SDS form an badly soluble Potassium salt.
May you can precipitate most of the SDS by adding for example Potassium Acetate.
This will work badly if you have a lot of Lithium in solution as I read (no experience with that). By the way, SDS is also badly soluble in the cold.
May cooling the sample closely to 0°C can also precipitate most of it.
However I don't know if this will also (co-) precipitate your protein or traces of it.
Furthermore Thermo offers detergent removal columns. They are filled with (I guess) Polystyrene beads, which are supposed to adsorb the deteregent on its surface. I must admit I never worked with it. But may this helps you:
https://www.thermofisher.com/order/catalog/product/87777
They also offer a HiPPR (better protein recovery).
Good luck
Hi, did you ever try that kit? 2-D Clean-Up Kit from GE ? I used it quite a bit and it works nicely for 2 -D gel sample were you must not have SDS. I used it for low protein concentration and was satisfied. I have to say I am not a fan of kits, but this is certainly one I would recommend.
Good luck
Here it comes: http://www.gelifesciences.com/webapp/wcs/stores/servlet/catalog/en/GELifeSciences-ch/products/AlternativeProductStructure_16720/80648451
Wolfgang
Did you try dialysis? it's a very old method, simple and very effective and no loss of protein!
There is a method for SDS removal from proteins using small scale ion-pair extraction described in the following paper:
Analytical Biochemistry
Volume 93, 1979, Pages 153-157
A micromethod for complete removal of dodecyl sulfate from proteins by ion-pair extraction
L.E.Henderson, S.Oroszlan, W.Konigsberg
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