M.T. Ardah Posted October 24, 2012 Share Posted October 24, 2012 Hi All, I am using Benzamidine sepharose beads to get rid of thrombin after GST tag cleavage. Afer I added the beads to the protein solution which contains Thrombin I took samples from the beads and the supernatant to check the purity of my protein in SDS and found that most of my protein bound to the beads. Any one have an aidea how to avoid this problem. Regards, Link to comment Share on other sites More sharing options...
CharonY Posted October 24, 2012 Share Posted October 24, 2012 This should not happen, unless your protein in question has an affinity to pABA. Did you use commercial columns? And if so, followed the manufacturer's instructions? Link to comment Share on other sites More sharing options...
M.T. Ardah Posted October 24, 2012 Author Share Posted October 24, 2012 Thax, Actually I am using in tubes not columns!! and what is pABA?? This should not happen, unless your protein in question has an affinity to pABA. Did you use commercial columns? And if so, followed the manufacturer's instructions? Link to comment Share on other sites More sharing options...
CharonY Posted October 24, 2012 Share Posted October 24, 2012 Sorry for the lab lingo, pABA is short for p-aminobenzamidine. It is more commonly linked to sepharose and my brain must have filled it in. Whether columns or tubes, it is still affinity purification. But it will depend on the precise protocol and material being used. Link to comment Share on other sites More sharing options...
Recommended Posts
Create an account or sign in to comment
You need to be a member in order to leave a comment
Create an account
Sign up for a new account in our community. It's easy!
Register a new accountSign in
Already have an account? Sign in here.
Sign In Now