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Is there posibility Reduced Glutathione cut GST-tag out of Protein


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#1 Hthongoc

Hthongoc

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Posted 19 April 2013 - 12:39 AM

I have tried to elute my GST-tag protein form Glutathione beads by 20 mM Reduced Glutathione in 50 mM Tris-HCl buffer pH 8.0.
I used one volume of elution buffer above with equal volume of Glutathione bead so the final concentration of GSH is 10mM.
I rotated the tube for 3 days. After that I found that about 40% of my protein lost GST-tag (on SDS-PAGE gel, I recorded 3 bands of GSt-tag protein, protein and GST).

I wonder if there is any possibility that GSH Glutathione cut GST-tag out of my target protein?
Have anyone here experienced it?
Or just I silly mistook to add Precission Protease into the tube?

Thank you for your help.




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