INDIGO Ni-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 INDIGO Ni-Agarose, based on high quality crosslinked agarose with 100 µm mean diameter, providing high flow rates in batch and FPLC applications. Using the novel, proprietary INDIGO ligand, purification of his-tagged proteins is possible in the presence of up to 20 mM DTT and 20 mM EDTA at a protein capacity considerably higher than for comparable competitor products. The novel INDIGO ligand cannot be stripped with EDTA, or cleaned at high pH using NaOH. However, without any regeneration, PureCube 100 INDIGO Ni-Agarose was re-used eight times without reduction in performance (Fig. 1). Cube Biotech offers PureCube 100 INDIGO Ni-Agarose, prepacked in affinity chromatography columns with 1 and 5 mL volume, ready to be used on automated liquid chromatography systems (e.g., FPLC).

Why PureCube 100 INDIGO Ni-Cartridges?

  • High flow rates AND dynamic binding capacity
  • Stable in 20 mM DTT and 20 mM EDTA, at pH 4-9
  • Compatible with eukaryotic cell culture media
  • Compatible with low pressure chromatography instruments
  • Two cartridge geometries: Standard and Compact

Indigo Cartridges in detail

Blau Chromatogramm 8 Läufe
Blau SDSPAGE 8 Läufe
Fig.1: PureCube 100 INDIGO Ni-Agarose can be re-used multiple times without regeneration. GFP was spiked into E.coli lysates and purified in eight aliquots on the same 1 ml column filled with PureCube 100 INDIGO Ni-Agarose. Between each run, the column was briefly washed with loading buffer containing PBS and 10 mM imidazole. No decrease in performance was observed, even after eight consecutive runs. Left: Chromatogram; Right: SDS-PAGE. M: Marker, L: Lysate, S: Lysate spiked with GFP.

Features

Feature 100 INDIGO NiCartridge, 1 mL 100 INDIGO NiCartridge, 5 mL
Usage Purification of his tagged proteins
Specifity Affinity to His-tagged proteins
Technology IMAC
Protein Binding capacity
80 mg (1ml cartridge)
400 mg (5ml cartridge)
Chelator stability 20 mM DTT; 20 mM EDTA
pH stability 4-9
Other Compatibilities 100% methanol, 100% ethanol, 8 M urea, 6 M guanidinium hydrochloride, 30% (v/v) acetonitrile
Bead size 50 -150 μm
Storage temperature 2-8 °C
Bead Ligand INDIGO 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 INDIGO Ni-Agarose, based on high quality crosslinked agarose with 100 µm mean diameter, providing high flow rates in batch and FPLC applications. Using the novel, proprietary INDIGO ligand, purification of his-tagged proteins is possible in the presence of up to 20 mM DTT and 20 mM EDTA at a protein capacity considerably higher than for comparable competitor products. The novel INDIGO ligand cannot be stripped with EDTA, or cleaned at high pH using NaOH. However, without any regeneration, PureCube 100 INDIGO Ni-Agarose was re-used eight times without reduction in performance (Fig. 1). Cube Biotech offers PureCube 100 INDIGO Ni-Agarose, prepacked in affinity chromatography columns with 1 and 5 mL volume, ready to be used on automated liquid chromatography systems (e.g., FPLC).

Why PureCube 100 INDIGO Ni-Cartridges?

  • High flow rates AND dynamic binding capacity
  • Stable in 20 mM DTT and 20 mM EDTA, at pH 4-9
  • Compatible with eukaryotic cell culture media
  • Compatible with low pressure chromatography instruments
  • Two cartridge geometries: Standard and Compact

Indigo Cartridges in detail

Blau Chromatogramm 8 Läufe
Blau SDSPAGE 8 Läufe
Fig.1: PureCube 100 INDIGO Ni-Agarose can be re-used multiple times without regeneration. GFP was spiked into E.coli lysates and purified in eight aliquots on the same 1 ml column filled with PureCube 100 INDIGO Ni-Agarose. Between each run, the column was briefly washed with loading buffer containing PBS and 10 mM imidazole. No decrease in performance was observed, even after eight consecutive runs. Left: Chromatogram; Right: SDS-PAGE. M: Marker, L: Lysate, S: Lysate spiked with GFP.

Features

Feature 100 INDIGO NiCartridge, 1 mL 100 INDIGO NiCartridge, 5 mL
Usage Purification of his tagged proteins
Specifity Affinity to His-tagged proteins
Technology IMAC
Protein Binding capacity
80 mg (1ml cartridge)
400 mg (5ml cartridge)
Chelator stability 20 mM DTT; 20 mM EDTA
pH stability 4-9
Other Compatibilities 100% methanol, 100% ethanol, 8 M urea, 6 M guanidinium hydrochloride, 30% (v/v) acetonitrile
Bead size 50 -150 μm
Storage temperature 2-8 °C
Bead Ligand INDIGO 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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INDIGO Ni-Cartridges 100
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PureCube 100 INDIGO Ni-Cartridge PureCube 100 INDIGO Ni-Cartridge
Cartridge for low pressure chromatography (FPLC), packed with 1 ml Agarose with 100 micron average bead size, equipped with a proprietary ligand for His affinity purification, loaded with nickel (II) sulfate For more information read...
Article number: 75301
Sales price: From €47.00 *
PureCube 100 Compact Cartridge Ni-INDIGO PureCube 100 Compact Cartridge Ni-INDIGO
Compact Cartridge for low pressure chromatography (FPLC), packed with 1 ml Agarose with 100 micron average bead size, equipped with a proprietary ligand for His affinity purification, loaded with nickel (II) sulfate
Article number: 75302
Sales price: From €49.00 *
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