Nickel NTA Chelating Superflow 4

Nickel NTA-Chelating Superflow 4 is an immobilized metal affinity chromatography (IMAC) resin that separates proteins via their differences in affinity to nickel. This product is recommended as a drop-in replacement for other tetra-dentate chelators.

Nickel NTA-Chelating Superflow 4 is generated from a proprietary tri-dentate chelating chemistry that has been optimized to provide increased flow rates (> 30 ml/min) and protein binding capacities (> 50 mg protein per ml resin). Along with no metal leakage properties, this resin is ideal for either large batch or column purification.

Cleanable, rechargeable and industrial process-ready, the Nickel NTA Chelating Superflow 4 resin is manufactured under cGMP and ISO 9001:2015 conditions. Regulatory Support Files are available as well as DMF with USFDA

The product is shipped pre-charged and ready to use. Optimizing the separation usually requires some method development.

The product is shipped pre-charged and ready to use. Optimizing the separation usually requires some method development.

Product Code: 937410SF4

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Table 1. Resin Characteristics

Bead Material Agarose
Bead Percentage 4%
Bead Size 60-160 µm
Crosslinking Highly Crosslinked Agarose (Superflow 4 Matrix)
Flow Rate 1 >23 mL/min (686 cm/h) @ 1.6 cm x 20cm column
Protein Binding Capacity>50 mg/mL
pH Stability 2-14
Storage Temperature 2-8oC
Storage Buffer 20% Ethanol
Metal Leaching Negligible Nickel Leaching (<0.5%)
Form Slurry
Chemical Stability 21mM EDTA, 20mM DTT, 1M NaOH, 6M Guanidine-HCl, 8M Urea. The use of primary amines (Tris, glycine, etc.) can weaken binding affinity and should not be utilized.
Physical Stability 2Negligible volume variation due to changes in pH or ionic strength.

1Linear flow rate = volumetric flow rate (cm3/h)/column cross-sectional area (cm2)

2Data refer to the coupled product, provided that the ligand can withstand the pH or chemical environment. Please note the following: pH stability, long term refers to the pH interval where the medium is stable over a long period of time without adverse effects on its subsequent chromatographic performance. pH stability, short term refers to the pH interval for regeneration and cleaning procedures.

Nickel NTA Chelating Superflow 4

Note: Chelating agents such as EDTA or citrate buffers can strip the Nickel ions from the resin. Avoid using these except for elution. The resin optimally binds at neutral to slightly basic pH (7-8).

Before you start:
1. Measure the required amount of suction-dried resin.
2. Wash the resin in a Buchner funnel or empty column with 5 bed volumes of DI water.
3. Remove the excess water by gentle suction and transfer the resin into a beaker or flask.
4. Determine Imidazole concentration for binding buffer, lower concentration gives higher yield but lower purity.

1. Storage Buffer: 20% Ethanol
2. Binding Buffer: 20 mM Sodium Phosphate buffer with 0.5 M NaCl, 20-40 mM Imidazole, pH 7-8
3. Alternative Binding Buffer: Tris-HCl (Tris may reduce binding affinity)
4. Elution Buffer: 20 mM Sodium Phosphate, 0.5 M NaCl, 500 mM (dependent on target protein) Imidazole
5. Alternative Elution Buffer: 10 mM EDTA

1. Wash the resin with at least 5 bed volumes of DI Water.
2. Equilibrate the washed resin in a column with 5-10 column volumes of binding buffer at 50-150 cm/h.
3. Load desired histidine-tagged protein sample onto the column and resin.
4. Pour binding buffer through the column and collect the flow-through. Measure absorbance of flow-through.
5. Elute with elution buffer using a stepwise or linear gradient and measure absorbance of eluent liquid.
6. Wash resin with 10-20 bed volumes of DI water to remove addition/unchelated metal ions.
7. Suction dry resin to remove residual water.
8. Place Nickel NTA Chelating Superflow 4 in 0.5 bed volumes of 20% ethanol.
9. Store resin at 2-8 °C.


Nickel NTA Chelating Superflow 4 (cat#937410SF4) is supplied in 20% ethanol. Store at 2-8 °C. Do not freeze. Do not expose the resin to acidic or basic conditions (unless it is prescribed in the instructions).
For technical service email or call (760) 929-0455

To Download Instructions for use:
Instructions for Use for Product Code: 937410SF4 Nickel NTA Chelating Superflow 4

Safety Data Sheets

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