Resin Equilibration and Binding and On Column TEV Cleavage
6:47
Packing and Eluting from Calmodulin Resin
8:20
Results: TAP Purification of U1 snRNP Complex in S. cerevisiae
9:55
Conclusion
Trascrizione
The overall goal of this protocol is to purify a native macromolecular complex of suitable quality for biophysical studies. This protocol serves not only as a way to purify a complex, but as a detailed guide to assessing the structural quality of
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The Tandem Affinity Purification (TAP) method has been used extensively to isolate native complexes from cellular extract, primarily eukaryotic, for proteomics. Here, we present a TAP method protocol optimized for purification of native complexes for structural studies.