HIS-Select^® Resins

• HIS-Select^® Nickel Affinity Gel
• HIS-Select^® High Flow (HF) Nickel Affinity Gel
• HIS-Select^® Cobalt Affinity Gel
• IMAC-Select Affinity Gel

This is the tool of choice for small to medium scale purification of histidine-tagged recombinant proteins. The graph below shows a typical purification using HIS-Select Nickel Affinity Gel. Lanes 7-10 demonstrate highly purified histidine-tagged protein, eluted with buffer plus 250 µm imidazole.

Features & benefits High Selectivity for Higher Purity Non-charged, hydrophilic linkage reduces non-specific binding One-step purification Capacity is greater than 15 mg/ml Minimal nickel leaching Support: 6% agarose Maximum flow rate: 150 cm/hr Maximum pressure: 5 psi Bead size: 45-165 µm

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HIS-Select HF (High Flow-Rate) Nickel Affinity Gel brings the superior selectivity of HIS-Select technology to a highly cross-linked agarose designed for higher flow rates and mechanical stability under pressure. This HIS-select product is designed for production scale purification and FPLC applications. As with other HIS-Select products, the non-charged, hydrophilic linkage of the NTA chelate group to the agarose ensures true one-step purification.

Features & benefits High Selectivity for Higher Purity Non-charged, hydrophilic linkage reduces non-specific binding One-step purification Capacity is greater than 15 mg/ml Minimal nickel leaching Support: highly cross-linked 6% beaded agarose Maximum flow rate: 3,000 cm/hr Maximum pressure: 200 psi Bead size: 45-165 µm

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For those who prefer to use cobalt as the affinity metal ion for HIS-tagged protein purification, our new HIS-Select Cobalt Affinity Gel allows for high purity, low non-specific binding and high binding capacity for your protein. HIS-Select Cobalt Affinity Gel also works well for purification of HIS-tagged proteins in native, denaturing, or mild reducing conditions.

Lane 1 – Sigma Wide Range Marker (M4038) Lane 2 – Empty Lane 3 – HIS-Select Cobalt Affinity Gel (H8162) Elution Lane 4 – Competitor C Cobalt Metal Affinity Resin Elution E.coli cell extract containing a HIS-tagged protein was purified using HIS-Select Cobalt Affinity Gel (H8162) and a leading competitor’s Cobalt Metal Affinity Resin using standard protocols for each. Samples were run on a 4-20% Tris-Glycine gel and stained with EZ-Blue Gel Staining Reagent (G1041).

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IMAC-Select Affinity Gel is an immobilized metal affinity chromatography (IMAC) product that is supplied as a metal-free gel allowing researchers to add a chelating metal of choice. IMAC-Select Affinity Gel is durable and can capture proteins that have an affinity for various metal ions. Other immobilized metal ions, such as iron or gallium, may be used to purify phosphoproteins. While most commonly used for purifying proteins it can also be used for purifying peptides.

Features & benefits Supplied as metal-free Support 6% beaded agarose Non-charged, hydrophilic linkage reduces non-specific binding For protein or peptide purification Metal binding capacity of 10–30 µmole/mL of resin Maximum flow rate: 75 cm/hr Maximum pressure: 30 psi Bead size: 45–160 µm

A 28 kDa histidine-tagged recombinant protein was purified from E.coli lysates using IMAC-Select Affinity Gel charged with either copper or zinc. Lanes 3 and 8 are the recombinant protein. Lanes 4 and 9 are the E.coli lysates containing the target protein. Lanes 5 and 10 are the purified protein after IMAC. Molecular Weight Marker used is ColorBurst™ Marker (Product No. C1992).

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