Protein Interest Group

The PIG discussion today lead to some discussion on IMAC.

We use different IMAC resins for different expression hosts and modalities.

What are your preferred modes and resins for IMAC?
Ni-NTA Superflow, Agarose, Excel, HiTrap, HIS-Trap,...
Modes: Automated, Batch Bind, Direct Inject, Magnetic, ...

Do you use any washes or methods that improve purity from a single step?
High Salt is super helpful for some programs.
TX-114 on E coli derived materials reduces endotoxin A LOT. Highly recommended. You can use HIC afterwards to remove the detergent.

In our labs, we primarily use Nickel ProBond (Invitrogen/Thermo). We primarily utilize batch bind/elution for primary affinity capture (this includes small-scale plate-based expression/purification as well as multiliter volume prep scale). We primarily do gravity flow for Ni-reverse clean up of proteases and contaminants.