PROTEINDEX™ HiBond™ Ni-NTA Agarose

PROTEINDEX™ HiBond™ Ni-NTA Agarose utilizes 4% agarose as the matrix support, and is functionalized with the chelate ligand Nitilotriacetic Acid (NTA), which is charged with nickel ions (Ni²⁺) to form a very stable octahedral structure with the nickel ion in the center. The structure of Ni-NTA complex is compatible with solutions that contain certain concentration of reducing agents, denaturing agents, detergents and other additives.

PROTEINDEX™ HiBond™ Ni-NTA Agarose can be used to purify 6xHis-tagged proteins expressed in various hosts, such as E.coli., yeast, insect cells and mammalian cells.

 

Product Highlights

• Suited for routine purification of histidine-tagged proteins
• High yield (typically > 40 mg protein /mL resins)
• Low leakage of nickel ions. Can be re-used for up to 8 preps*

*Note: No significant decrease in yield after 8 repetitive uses of resin when recommended buffers are used. Refer to Product Details for compatible reagents.

 

Applications

• Manual batch or gravity flow purification
• Routine His tag protein purification for structural biology and functional assays
• Protein-protein and protein-nucleic acid interaction studies

 

Tech Notes

One-step purification using ProteIndex™ HiBond™ Ni-NTA Agarose 6 Fast Flow resins

Figure 1. E.coli lysate containing a 50 kDa target protein was directly loaded on HiBond™ Ni-NTA Agarose 6 FF column. The column was then washed sequentially with 0 mM, 5 mM, and 20 mM imidazole solution before elution with 300 mM imidazole. The target protein was analyzed by SDS-PAGE. CL: raw lysate; FT: unbound flow-through; W1, W2 and W3: washes; E1 and E2: eluate fractions.

Product Details

Ligand	Nickel
Binding capacity	>40 mg 6xHis-tagged protein/mL medium
Matrix	4% agarose supplied as 50% slurry
Particle size	45 μm - 165 μm
Maximum pressure	0.1 MPa (1 bar)
Recommended flow rate	Gravity flow, <75 cm/hour
Chemical compatibility	Reducing agents 5 mM DTE 0.5-1mM DTT 20 mM β-mercaptoethanol 5 mM TCEP 10 mM reduced glutathione (GSH) Denaturing agents 8 M urea, 6 M Gua-HCl (Tested for one week at 40°C) Detergents 2% Triton™ X-100 (nonionic) 2% Tween™ 20 (nonionic) 2% NP-40 (nonionic) 2% cholate (anionic) 1% CHAPS (zwitterionic) Other additives 500 mM imidazole 20% ethanol 50% glycerol 100 mM Na2SO4 1.5 M NaCl 1 mM EDTA 60 mM citrate Commonly used buffer 50 mM sodium phosphate, pH 7.4 100 mM Tris-HCl, pH 7.4 100 mM Tris-acetate, pH 7.4 100 mM HEPES, pH 7.4 100 mM MOPS, pH 7.4 100 mM sodium acetate, pH 4
Storage temperature	2 - 8°C
Storage buffer	1x PBS containing 20% ethanol

 

Product is shipped at ambient temperature. Upon receipt store at 2 - 8°C. Do not Freeze.

      

 



 

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Marvelgent Biosciences Inc.

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