After clevage of my protein fusion GST, My protein is cleaved but it is still stocked with beads. The protein is not eluted in the buffer of clevage.
Do you have any suggestions?
Thanks
Resolution:
Only so many possibilities - either it has become insoluble due to the lack of its GST partner, has been hydrophobically or ionically modified so that it now interacts with the column (and needs to be eluted) or the cleavage did not work like you think it did and the protein is still associated with its GST partner. These are the issues we confront regularly in our custom thrombin cleavage service.
We would be glad to take care of the problem for you. If interested, go to http://www.virusys.com/Services/ag103.html and request a quotation.
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