His-taged protein is in 8M Urea, how to get rid of urea? - Ni-NTA column purified protein in 8M urea buffer (May/18/2005 )
Hello! I have purified my protein through Ni-NTA column , Now it is in Elution buffer containing 8M urea, I want to use this protein for ELISA. For this I need your guidance on How to get rid of urea , and other precautions that may help me!
-mh450-
dear researcher,
it is very easy for u to get rid of the Urea from solution. just dialyse it with 2 or 3 litre of the same solution in which u have yr enzyme. like if u have it in PBS or other buffer. after 2 to 5 Hrs of Dialysis u will get rid of that.
regards
BS
-singh-
Desalting with sephadesx G25 columns produces the same effect at much shorter time
-sharath-
QUOTE (sharath @ May 31 2005, 04:00 PM)
Desalting with sephadesx G25 columns produces the same effect at much shorter time
ght
Hi,
is using the sephadex G25 column better than gradually reducing the UREA concentration and eventually removing it completely?...cos when i tried dialysing straight into PBS, my protein (40% hydrophobic) precipitated! I currently use the Pierce dialysis cassette.....thank you for any insight
-ggUss-