His-Tag Purification
This protocol will only work if you’ve included a specific 6-12 repeating Histidine sequence in the amino acid code of the expressed protein of interest. You should be adding the sequence at the N-Terminal or C-Terminal, which is better will depend on the protein structure and should be decided based upon computational modelling to determine which end sticks out more or through trial-and-error experimentation.
Our price modelling has shown that Nickel-Agarose purification is the primary scaling cost of protein production. Finding cheap alternatives to this protocol is of paramount importance.
Protocol: Filter & Centrifuge His-Tag Purification
If you don’t have $20,000 to throw down on a second-hand FPLC, this protocol is useful for small scale purification of protein. It isn’t particularly scaleable
Protocol: Column Chromatography His-Tag Purification (FPLC)
Industrial standard for most scaled biomolecule purification. Ungodly expensive, oligopoly market. Chief contributor to scaling expense of production.