PhyNexus IMAC (Immobilized Metal Affinity Chromatography) PhyTip® columns purify or immobilize functional, recombinant His tagged protein or antibody quickly at high purity and concentration. IMAC separation resin in pipette tips is used to generate highly enriched sample from microliter to milliliter volumes in a reproducible, automatable format. The unique PhyTip column design allows bidirectional flow of the sample through the column to increase capture and elution of the target protein, maximizing the use of the resin and leading to the highest yield possible.

  • IMAC (Ni-IMAC) affinity columns routinely produce samples of greater than 90% purity. The purity is demonstrated by the absence of background proteins.
  • Recover 60-80% of fully functional protein with greater than 90% purity
  • Process small sample volumes in a reproducible, high throughput automation
  • IMAC(Ni-IMAC) tips are processed in a high throughput automation
  • Elution volumes as low as 10 μL, producing enrichment factors as high as 50 fold, with concentrations of purified protein of up to 10 mg/mL

IMAC columns with our proprietary Dual Flow Chromatography provide complete antibody binding and high final concentrations. PhyTip columns are compatible with most 8, 12, and 96-channel automation liquid handling robots including Agilent TechnologiesBeckman CoulterDynamic DevicesPerkin ElmerHamilton, and Tecan.

PhyTip columns are unique capture, purification and enrichment tools from PhyNexus designed for micro volume protein sample preparation. PhyTip Columns are available for a variety of liquid handling platforms and contain specific affinity resins for application specific requirements. Samples for purification and enrichment must be clear and free from particulate matter. It is highly recommended to centrifuge samples and use the clear supernatant only, prior to use with PhyTip columns.

200+ PhyTip columns with IMAC resin:

For a 200 µL sample with 5 µg His-Tagged Fab containing 1 mg BSA processed using the conditions shown below, greater than 50% of the original HisTagged Fab mass is recovered in the final sample volume. In addition, this recovered His-Tagged is purified to over >95% purity as determined by SDSPAGE with Coomassie detection.

Capture: 200 µL sample captured by passing through the resin bed for two cycles at a flow rate of 250 uL per minute.

Purify: 200 µL of PhyNexus IMAC Wash Buffer I, passed over the resin bed for two cycles at a flow rate of 500 µL/min followed by a second wash with same buffer, passed over the resin bed for two cycles at a flow rate of 500 µL/min.

Enrich: elute the protein into solution with 10µL of IMAC Elution Buffer, passed over the resin bed for five cycles at a flow rate of 500 µL/min.

1000+ PhyTip columns with IMAC resin:

For a 500 µL sample with 5 µg His-Tagged Fab containing 1 mg BSA, processed using the conditions shown below, greater than 50% of the original HisTagged Fab mass is recovered in the final sample volume. In addition, this recovered His-Tagged Fab is purified to over >95% purity as determined by SDSPAGE with Coomassie detection.

Capture: 500 µL sample captured by passing through the resin bed for two cycles at a flow rate of 250 uL per minute.

Purify: 1000 µL of PhyNexus IMAC Wash Buffer I, passed over the resin bed for two cycles at a flow rate 500 µL/min followed by a second wash with the same buffer, passed over the resin bed for two cycles at a flow rate 500 µL/min.

Enrich: elute the protein into solution with 15µL of PhyNexus IMAC Elution Buffer, passed over the resin bed for five cycles at a flow rate of 500 µL/min.

Select your resin, robotic system, tip and resin volume and see the results in the table below.

IMAC Columns