NTA-Chelating Superflow 4

NTA-Chelating Superflow 4 is an immobilized metal affinity chromatography (IMAC) resin that separates proteins via their differences in affinity to a variety of metal ions (i.e. Ni, Fe, Co, Cu, or Zn).

NTA-Chelating Superflow 4 is a tetra-dentate chelator that typically has high binding capacities in comparison to other chelators. It is shipped uncharged so that the customer can select which transition metal to chelate with the resin. Typically, users find Ni, Fe, Co, Cu, or Zn to be the most effective. Optimizing the separation usually requires some method development.
NTA-Chelating Superflow 4 is compatible with a variety of chemicals, pH values and standard clean-in-place procedures. It is stable through multiple cycles of cleaning and reuse. This product is recommended for the purification of polyhistidine-tagged proteins as it provides higher selectivity and no metal leaching.

Product Code: 937422SF4
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Table 1. Resin Characteristics

Bead Material Agarose
Bead Percentage 4%
Bead Size60-160 µm
Binding Capacity (Hb) >50 mg/mL
Flow Rate1>30 mL/min (>1020 cm/hr) @ 25oC
pH Stability22-9
Storage Temperature2-8o C
Storage Buffer 20% Ethanol
FormSlurry
Chemical Stability2Stable in commonly used aqueous buffers, salts, detergents, and reducing agents. Compatible with <8M Urea, <6M Guanidine HCl, and 1M NaOH.
Physical Stability2Negligible 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.

Instructions for Use

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.

Charging NTA Chelating Superflow 4 with Divalent (M2+) metal ions
1. Prepare a 75mg/mL solution of the desired Divalent metal ion (M2+, M = Zn, Ni, Co, Cu, etc) in DI
water.
2. Mix until all the metal ion solution is dissolved (~5 to 10 minutes).
3. Pour solution into vessel containing NTA Chelating Superflow 4 resin.
4. Allow mixture to shake/rock for 1 hour at room temperature.
5. After 1 hour, the coordination of the metal ion to the NTA Chelating Superflow 4 resin is
complete. Proceed to pour the charged resin into a Buchner funnel and suction dry.
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 metal coordinated NTA Chelating Superflow 4 in 0.5 bed volume of 20% ethanol.
9. Store in 2-8oC.
NOTE: Although the use of sulfate counter ions (SO42-) is the most used form, other counter ions such as: Cl and NO3 are also acceptable.

Storage
Uncoordinated NTA Chelating Superflow 4 (cat#37410) is supplied in 20% ethanol. Store at 2-8oC. Do not
freeze. Do not expose the resin to acidic or basic conditions (unless it is prescribed in the instructions).

To Download Instructions for use:
Instructions for Use 937422SF4 NTA Chelating Superflow 4

Safety Data Sheets

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