MICR 302 Lecture Notes - Lecture 6: Protein A, Immunoglobulin G, Zygosity
7 Apr 2018
School
Department
Course
Professor
Document Summary
Tap tags for puri cation of protein complexes help to identify protein-protein interactions: Use sequential a nity puri cation rather than singular a nity puri cation: Load the puri ed protein extract from tap tagged yeast cells into a column that contains agarose/separose beads that are igg coated. The tap tag has 2 parts that are separated by a cleavage site that can both be baited by the beads. The protein a part of the tap tag will bind to the igg beads, and the remaining proteins are washed from the column. Add tev + bu er to cleave the site between protein a and cbp (calmodulin binding protein), liberating the protein of interest (bait protein) Load the elution from the rst column into a column containing. Calmodulin coated beads that will only bind cbp in the presence of ca2+. The protein of interest will be captured again, and any other non-speci c protein will be washed.
