Purification of Native Complexes for Structural Study Using a Tandem Affinity Tag Method

385 مشاهدة

استشهد بها 4

10:36 دقيقة

يوليو 27, 2016

10.3791/54389-v

يوليو 27, 2016

385 مشاهدة
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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.

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Tandem Affinity Purification

Chapters in this video

0:05

Title

0:38

Harvesting S. cerevisiae Cells

1:44

Lyse Cells and Lysate Clarification

4:43

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

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