Binding of His-tagged Proteins to the HisPur Cobalt Beads
4:39
Binding of the Peptide to the His-tagged Protein Immobilized on the Beads
5:59
Elution from Cobalt Beads
7:07
Electrophoresis, Western Blotting, and Analysis of Results
7:58
Results: Use of Immunoblot Assay to Study the Interaction between Sortilin Immobilized on Cobalt Beads and myc-tagged Peptide
9:18
Conclusion
Transcript
The overall goal of this procedure is to detect detergent-sensitive interactions between membrane proteins or other proteins. This procedure requires one protein to be histidine-tagged and over expressed in the cells, while the other is a myc-tagg
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We describe a protocol for detection of detergent-sensitive interactions between membrane proteins using binding of the sorting receptor, sortilin, to the first luminal loop of the glucose transporter protein, GLUT4, as an example.
