Ni-NTA Cartridges 100


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 introduces the novel PureCube 100 Ni-NTA Agarose, based on high quality crosslinked agarose with 100 µm mean diameter, providing high flow rates in batch and FPLC applications. Due to the excellent flow properties of PureCube 100 Agarose, combined with the high capacity NTA ligand, high dynamic yield and protein purity can be achieved. Even at flow rates of 7 ml/min, more than 20 mg/ml his-tagged protein could be purified (Fig.1). PureCube 100 Ni-NTA Agarose shows high affinity to his-tagged proteins. Recovery rates of 85-100% were achieved, even with low concentrated protein samples (Fig.2). Cube Biotech offers PureCube 100 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 100 Ni-NTA Cartridges provide:


PureCube 100 Ni-NTA in detail

Dynamic Diagramm_mL R
Dynamic binding SDS R
Fig. 1: High dynamic binding capacity of PureCube 100 Ni-NTA Agarose. GFP was spiked into E.coli lysates and purified on a 1 mL PureCube Cartridge filled with PureCube 100 Ni-NTA Agarose at flow rates from 1 to 7 ml/min, and in a gravity flow batch procedure. Left: Flow diagram, Right: SDS-PAGE analysis of elution fractions.
Protein Recovery;n Recovery_0916_2_R
Fig.2: Excellent protein recovery with PureCube 100 Ni-NTA Agarose. GFP was spiked into E.coli lysates in different quantities and purified on a 1 mL PureCube Cartridge filled with PureCube 100 Ni-NTA Agarose. Recovered GFP was quantified by absorption at 488 nm.

Features

Feature PureCube 100 Ni-NTA Cartridge PureCube 100 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 ~100 μm
Storage temperature 2-8 °C
Bead Ligand 100 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 6% 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.
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 introduces the novel PureCube 100 Ni-NTA Agarose, based on high quality crosslinked agarose with 100 µm mean diameter, providing high flow rates in batch and FPLC applications. Due to the excellent flow properties of PureCube 100 Agarose, combined with the high capacity NTA ligand, high dynamic yield and protein purity can be achieved. Even at flow rates of 7 ml/min, more than 20 mg/ml his-tagged protein could be purified (Fig.1). PureCube 100 Ni-NTA Agarose shows high affinity to his-tagged proteins. Recovery rates of 85-100% were achieved, even with low concentrated protein samples (Fig.2). Cube Biotech offers PureCube 100 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 100 Ni-NTA Cartridges provide:


PureCube 100 Ni-NTA in detail

Dynamic Diagramm_mL R
Dynamic binding SDS R
Fig. 1: High dynamic binding capacity of PureCube 100 Ni-NTA Agarose. GFP was spiked into E.coli lysates and purified on a 1 mL PureCube Cartridge filled with PureCube 100 Ni-NTA Agarose at flow rates from 1 to 7 ml/min, and in a gravity flow batch procedure. Left: Flow diagram, Right: SDS-PAGE analysis of elution fractions.
Protein Recovery;n Recovery_0916_2_R
Fig.2: Excellent protein recovery with PureCube 100 Ni-NTA Agarose. GFP was spiked into E.coli lysates in different quantities and purified on a 1 mL PureCube Cartridge filled with PureCube 100 Ni-NTA Agarose. Recovered GFP was quantified by absorption at 488 nm.

Features

Feature PureCube 100 Ni-NTA Cartridge PureCube 100 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 ~100 μm
Storage temperature 2-8 °C
Bead Ligand 100 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 6% 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.
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Ni-NTA Cartridges 100
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PureCube 100 Ni-NTA Cartridge PureCube 100 Ni-NTA Cartridge
Cartridge for low pressure chromatography (FPLC), packed with 1 ml NTA Agarose with 100 micron average bead size, loaded with nickel (II) sulfate
Article number: 74301
Sales price: From €47.00 *
PureCube 100 Compact Cartridge Ni-NTA PureCube 100 Compact Cartridge Ni-NTA
Compact Cartridge for low pressure chromatography (FPLC), packed with 1 ml NTA Agarose with 100 micron average bead size, loaded with nickel (II) sulfate
Article number: 74302
Sales price: From €47.00 *
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