We routinely perform E.coli expression and purification services for our customers using our AKTA Explorers and AKTA Xpress 4-pack. Our standard purification protocol uses nickel purification of a target protein that contains a Tev-protease cleavable His6 tag. A “subtractive” or “ortho” nickel purification is performed after TEV cleavage of the His6 tag as shown in the diagram below.
Depending on the purity of the target protein at this point, we may perform an ion exchange purification step before performing a gel filtration step. The target protein’s concentration, buffer conditions, and aliquot size are adjusted to meet your needs.
We perform all of the above steps for a charge of $777. We generally try several expression conditions and will scout column requirements to achieve adequate purity of your protein. Our goal is to provide you with our best attempt at purifying your protein from a 2L E. coli expression. You will receive from us your protein as well as detailed documentation of the protein’s preparation. If the yield meets your needs, then you are all set, otherwise we are happy to quote you a price for perform additional preps.
We can also tackle protein purifications that do not involve nickel affinity. Our general charge of $777 is good for any three column purification.
Please contact us for more information.