Ni-NTA Cartridges


Agarose guide
The His tag is the most widely used affinity tag due to its small size, low immunogenicity, and versatility under native or denaturing conditions, as well as in presence of detergents and many other additives. Cube Biotech offers high-performance Ni-NTA Agarose, prepacked in affinity chromatography columns with 1 and 5 mL volume, ready to be used on automated liquid chromatography systems (e.g., FPLC).

PureCube Ni-NTA Cartridges provide:

Ni-NTA cartridges from Cube Biotech was successfully used in the following publications:


Ni-NTA in detail

Dynamic Diagramm_mL R
Dynamic binding SDS R
Fig. 1: High dynamic binding capacity of PureCube Ni-NTA Agarose. GFP was spiked into E.coli lysates and purified on a 1 mL PureCube Cartridge filled with PureCube Ni-NTA Agarose. Different flow rates were tested too see how good each performes, for this purpose flow rates between 1 mL/min up tp 7 ml/min were choosen. Left: Flow rate compairing diagram, Right: SDS-PAGE comparing the elution factions.
Protein Recovery;n Recovery_0916_2_R
Fig.2: PureCube Ni-NTA Agarose provided an ecxellent elution rate. GFP was spiked into E.coli lysates in different quantities and purified on a 1 mL PureCube Cartridge filled with PureCube Ni-NTA Agarose. Recovered GFP was quantified by absorption at 488 nm.

Features

Feature Ni-NTA Cartridge Ni-NTA COMPACT Cartridge
Usage Purification of his tagged proteins
Specifity Affinity to His-tagged proteins
Protein Binding capacity
80 mg (1ml cartridge)
400 mg (5ml cartridge)
Chelator stability Stable in buffer containing 10 mM DTT and 1 mM EDTA
pH stability 2-14
Other Compatibilities 100% methanol, 100% ethanol, 8 M urea, 6 M guanidinium hydrochloride, 30% (v/v) acetonitrile
Bead size 40 μm
Storage temperature 2-8 °C
Bead Ligand Ni-NTA
Bed Volume 1 ml or 5 ml
Dimensions of cartridges
6.2mm X 50 mm (1ml cartridge)
11mm X 80 mm (5ml Cartridge)
5mm X 35 mm (1ml cartridge)
17mm X 35mm (5ml cartridge)
Column Body Material
Polypropylene (1ml cartridge)
Acrylate (5ml cartridge)
Polypropylene
Inlet 10-32 UNF female thread
Outlet 10-32 UNF female thread
Matrix 7.5% highly cross-linked agarose
Max. Flow Rate 6 mL/min
Recommended Flow Rate 0.5-2.0 mL/min
Recommended Operational Pressure
Up to 5 bar (72 psi) (1ml cartridge)
Up to 3 bar (42 psi) (5ml cartridge)
Required Buffers for purification under native conditions
 
  • Native Lysis buffer: NaH2PO4 50 mM, NaCl 300 mM, Imidazole 10 mM, pH 8 Optional: Add 1 tablet protease inhibitor cocktail to the Lysis Buffer. Tip: Lysis Buffer contains 10 mM imidazole to prevent binding of untagged proteins. If His-tagged proteins do not bind under these conditions, reduce the imidazole concentration to 1-5 mM.
  • Native Wash buffer: NaH2PO4 50 mM, NaCl 300 mM, Imidazole 20 mM, pH 8
  • Native Elution buffer: NaH2PO4 50 mM, NaCl 300 mM, Imidazole 250- 500 mM, pH 8
Required Buffers for purification under denaturing conditions
 
  • Denaturing Lysis buffer: NaH2PO4 100 mM, Tris base 10 mM, Urea 8 M, pH 8.0, Optional: Benzonase® nuclease (e.g. Merck Milipore, #707464)
  • Denaturing Wash buffer: NaH2PO4 100 mM, Tris base 10 mM, Urea 8 M, pH 6.3NaH2PO4 100 mM
  • Denaturing Elution buffer: NaH2PO4 100 mM, Tris base 10 mM, Urea 8 M, pH 4.5 Tip: If the target protein is acid-labile, elution can be performed with 250-500 mM imidazole. Note: Due to urea dissociation, adjust the pH immediately before use.

References

1. Stressler, T. et al. (2016). A Novel Glutamyl (Aspartyl)-Specific Aminopeptidase A from Lactobacillus delbrueckii with Promising Properties for Application. PLOS ONE. 11. e0152139. 10.1371/journal.pone.0152139.
2. Yang, M. et al. (2019). Rational Design of Alginate Lyase from Microbulbifer sp. Q7 to Improve Thermal Stability. Marine Drugs. 17. 378. 10.3390/md17060378.
3. Ewert, J. et al. (2017). Influence of the metal ion on the enzyme activity and kinetics of PepA from Lactobacillus delbrueckii. Enzyme and Microbial Technology. 110. 10.1016/j.enzmictec.2017.10.002.
4. Stressler, T. et al. (2018). A natural variant of arylsulfatase from Kluyveromyces lactis shows no formylglycine modification and has no enzyme activity. Applied Microbiology and Biotechnology. 102. 10.1007/s00253-018-8828-5.
5. Yang, M. et al.(2019). Study on expression and action mode of recombinant alginate lyases based on conserved domains reconstruction. Applied Microbiology and Biotechnology. 103. 10.1007/s00253-018-9502-7.
Agarose guide
The His tag is the most widely used affinity tag due to its small size, low immunogenicity, and versatility under native or denaturing conditions, as well as in presence of detergents and many other additives. Cube Biotech offers high-performance Ni-NTA Agarose, prepacked in affinity chromatography columns with 1 and 5 mL volume, ready to be used on automated liquid chromatography systems (e.g., FPLC).

PureCube Ni-NTA Cartridges provide:

Ni-NTA cartridges from Cube Biotech was successfully used in the following publications:


Ni-NTA in detail

Dynamic Diagramm_mL R
Dynamic binding SDS R
Fig. 1: High dynamic binding capacity of PureCube Ni-NTA Agarose. GFP was spiked into E.coli lysates and purified on a 1 mL PureCube Cartridge filled with PureCube Ni-NTA Agarose. Different flow rates were tested too see how good each performes, for this purpose flow rates between 1 mL/min up tp 7 ml/min were choosen. Left: Flow rate compairing diagram, Right: SDS-PAGE comparing the elution factions.
Protein Recovery;n Recovery_0916_2_R
Fig.2: PureCube Ni-NTA Agarose provided an ecxellent elution rate. GFP was spiked into E.coli lysates in different quantities and purified on a 1 mL PureCube Cartridge filled with PureCube Ni-NTA Agarose. Recovered GFP was quantified by absorption at 488 nm.

Features

Feature Ni-NTA Cartridge Ni-NTA COMPACT Cartridge
Usage Purification of his tagged proteins
Specifity Affinity to His-tagged proteins
Protein Binding capacity
80 mg (1ml cartridge)
400 mg (5ml cartridge)
Chelator stability Stable in buffer containing 10 mM DTT and 1 mM EDTA
pH stability 2-14
Other Compatibilities 100% methanol, 100% ethanol, 8 M urea, 6 M guanidinium hydrochloride, 30% (v/v) acetonitrile
Bead size 40 μm
Storage temperature 2-8 °C
Bead Ligand Ni-NTA
Bed Volume 1 ml or 5 ml
Dimensions of cartridges
6.2mm X 50 mm (1ml cartridge)
11mm X 80 mm (5ml Cartridge)
5mm X 35 mm (1ml cartridge)
17mm X 35mm (5ml cartridge)
Column Body Material
Polypropylene (1ml cartridge)
Acrylate (5ml cartridge)
Polypropylene
Inlet 10-32 UNF female thread
Outlet 10-32 UNF female thread
Matrix 7.5% highly cross-linked agarose
Max. Flow Rate 6 mL/min
Recommended Flow Rate 0.5-2.0 mL/min
Recommended Operational Pressure
Up to 5 bar (72 psi) (1ml cartridge)
Up to 3 bar (42 psi) (5ml cartridge)
Required Buffers for purification under native conditions
 
  • Native Lysis buffer: NaH2PO4 50 mM, NaCl 300 mM, Imidazole 10 mM, pH 8 Optional: Add 1 tablet protease inhibitor cocktail to the Lysis Buffer. Tip: Lysis Buffer contains 10 mM imidazole to prevent binding of untagged proteins. If His-tagged proteins do not bind under these conditions, reduce the imidazole concentration to 1-5 mM.
  • Native Wash buffer: NaH2PO4 50 mM, NaCl 300 mM, Imidazole 20 mM, pH 8
  • Native Elution buffer: NaH2PO4 50 mM, NaCl 300 mM, Imidazole 250- 500 mM, pH 8
Required Buffers for purification under denaturing conditions
 
  • Denaturing Lysis buffer: NaH2PO4 100 mM, Tris base 10 mM, Urea 8 M, pH 8.0, Optional: Benzonase® nuclease (e.g. Merck Milipore, #707464)
  • Denaturing Wash buffer: NaH2PO4 100 mM, Tris base 10 mM, Urea 8 M, pH 6.3NaH2PO4 100 mM
  • Denaturing Elution buffer: NaH2PO4 100 mM, Tris base 10 mM, Urea 8 M, pH 4.5 Tip: If the target protein is acid-labile, elution can be performed with 250-500 mM imidazole. Note: Due to urea dissociation, adjust the pH immediately before use.

References

1. Stressler, T. et al. (2016). A Novel Glutamyl (Aspartyl)-Specific Aminopeptidase A from Lactobacillus delbrueckii with Promising Properties for Application. PLOS ONE. 11. e0152139. 10.1371/journal.pone.0152139.
2. Yang, M. et al. (2019). Rational Design of Alginate Lyase from Microbulbifer sp. Q7 to Improve Thermal Stability. Marine Drugs. 17. 378. 10.3390/md17060378.
3. Ewert, J. et al. (2017). Influence of the metal ion on the enzyme activity and kinetics of PepA from Lactobacillus delbrueckii. Enzyme and Microbial Technology. 110. 10.1016/j.enzmictec.2017.10.002.
4. Stressler, T. et al. (2018). A natural variant of arylsulfatase from Kluyveromyces lactis shows no formylglycine modification and has no enzyme activity. Applied Microbiology and Biotechnology. 102. 10.1007/s00253-018-8828-5.
5. Yang, M. et al.(2019). Study on expression and action mode of recombinant alginate lyases based on conserved domains reconstruction. Applied Microbiology and Biotechnology. 103. 10.1007/s00253-018-9502-7.
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Ni-NTA Cartridges
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PureCube Compact Cartridge Ni-NTA PureCube Compact Cartridge Ni-NTA
Compact Cartridge for low pressure chromatography (FPLC), packed with 1 ml NTA Agarose, loaded with nickel (II) sulfate
Article number: 31302
Sales price: From €33.00 *
PureCube Ni-NTA Cartridge PureCube Ni-NTA Cartridge
Cartridge for low pressure chromatography (FPLC), packed with 1 ml NTA Agarose, loaded with nickel (II) sulfate
Article number: 31301
Sales price: From €33.00 *
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