IMAC PhyTip® Columns used for automated Purification of Histidine Tagged Proteins.

The histidine tag is the most commonly used tag for isolation of recombinant proteins. This tag typically consist of six or more histidine amino acids. Immobilized metal affinity chromatography (IMAC) separation resins are used for purification of his-tagged proteins. Elution can be achieved by mild elution with buffer containing imidazole (100 to 250 mM).

IMAC PhyTip columns are packed with high capacity Nickel-IMAC affinity resin and are used to purify biologically functional, recombinant His-tagged recombinant proteins and consistently generate purified samples of high purity and concentration by dual flow chromatography. The optimal resin bed volume of the column depends on the total amount of protein present in your samples. If you are working with very low protein concentrations of 10 μg/ml or lower, we recommend using a resin bed volume one or two sizes bigger than indicated below for an effective purification process.

PhyTip columns purified samples by repeated back-and-forth cycles - a process called dual flow chromatography - allowing complete binding interactions in each purification step.

IMAC PhyTip columns are packed with a high capacity IMAC resin, which routinely generates purified samples of his-tagged protein with 60 to 80% sample recovery. These columns routinely produce his-tagged protein samples of >90% purity as measured by the absence of background proteins present in the purified sample.

Unique column design with a packed resin bed results in binding of the entire sample to a small resin bed volume. The purified sample can be eluted in small volumes, resulting in a highly concentrated sample. PhyTip columns process samples by dual flow chromatography which is a gentle purification process for high levels of biologically active protein. Protein A PhyTip columns comes with dual flow chromatography protocols that have be validated with real protein and can be used on any automated liquid handling system.