Gloria,
Here is how we are now doing it, courtesy of Yu-Jing's and Merlin's fine development work. We have also tried it to quantitatively remove tags by TEV cleavage, but we haven't solved that problem yet.
Regards,
Michael
Tan,
Yu-Jing, Wei-Han Wang, Yi Zheng, Jinlan Dong, Giovanni Stefano, Federica
Brandizzi, R. Michael Garavito, Gavin E. Reid, and Merlin L. Bruening (2012)
Limited Proteolysis Via Millisecond Digestions in Protease-Modified Membranes.
Analytical Chemistry 84(19), 8357-63
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R. Michael Garavito, Ph.D.
Professor of Biochemistry & Molecular Biology
Michigan State University
East Lansing, MI 48824-1319
Office: (517) 355-9724 Lab: (517) 353-9125
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On Oct 8, 2013, at 3:34 PM, Gloria Borgstahl wrote:
We are trying to apply this approach for the first time to define a crystallizable domain of our protein of interest. Can anyone send a protocol that includes exactly how to do the mass spectrometry measurement? Our core lab here doesn't know and I'm just not that gifted. Happy October, G