Un abonnement à JoVE est nécessaire pour voir ce contenu. Connectez-vous ou commencez votre essai gratuit.
Chapitres
- 00:05Titre
- 00:52Collecting the Cells
- 01:39Lysing the Cells
- 02:32Equilibrating the STREP Beads
- 03:56STREP Affinity Purification (AP)
- 06:01Equilibrating the FLAG Beads
- 07:04FLAG Immunoprecipitation (IP)
- 08:39Results: A Cyclin-CDK Complex is Successfully Purified by Tandem Affinity Purification (TAP)
- 10:26Conclusion
We describe here a novel, robust, and efficient tandem affinity purification (TAP) method for the expression, isolation, and characterization of protein complexes from eukaryotic cells. This protocol could be utilized for the biochemical characterization of discrete complexes as well as the identification of novel interactors and post-translational modifications that regulate their function.
