Dear Manjula Ramu, You might want to consider immobilizing your thrombin-containing mixture on a matrix (e.g. Bio-Rad's Affi-Gel, but there are many alternatives) and then use the slurry for cleaving your recombinant protein. That way, you never contaminate your sample with either thrombin or BSA.
Good luck, Pedro -- Pedro J. B. Pereira, PhD IBMC - Biomolecular Structure Group Rua do Campo Alegre, 823 4150-180 Porto Portugal Tel. +351 226 074 900 Fax +351 226 099 157 E-mail [email protected] From: Manjula Ramu <[email protected]> To: [email protected] Date: 10/08/2014 02:33 PM Subject: [ccp4bb] How to separate BSA present in Thrombin Hi all, My protein size is nearly the same size of BSA and it has His tag. If I have to cleave the tag I have to use Thrombin which is having huge amount of BSA in it. Now how will I separate this BSA from my protein. Thanks and Regards, Manjula R Research Scholar Department of Biophysics National Institute of Mental Health and Neurosciences Bengaluru-29, Karnataka INDIA E-mail: [email protected] Mobile no:+91-9538553356 http://www.nimhans.kar.nic.in/
