NTA Agarose


Agarose purification
The His tag is the most widely used affinity tag due to its small size and versatility under native and denaturing conditions, as well as in presence of detergents and many other additives. Cube Biotech offers high-performance PureCube NTA Agarose, based on BioWorks Workbeads. NTA Agarose can be loaded with transition metals to obtain different specificities for his-tagged proteins, or for other applications such as purification of phosphorylated proteins. NTA Agarose is also available as pre-packed cartridge for FPLC applications. For purification from diluted solutions, NTA MagBeads are recommended. Do you prefer IDA? We offer the same high-quality resin with that chelating ligand! (what's the difference?)

Why PureCube NTA Agarose?

PureCube NTA Agarose from Cube Biotech was successfully used in the following publications:

 Coupled molecule / metal ionPurified substanceYearAuthor
Gallium Phages 2019 Schönberger N., Braun R., Matys S., Lederer F-L., Lehmann F., Flemming K., Pollmann K.1

Features

Usage Unloaded NTA Agarose for costumized purpose
Specifity Depending of loaded metal ion (E.g. Nickel for His-affinity)
Binding capacity Depending on loaded metal Ion
Stability Stable in pH2-14; 100% methanol, 100% Ethanol, 8M urea, 6M Guanidinium hydrochloride, 30% acetonitrile
Particle diameter ~40 µm
Filling quantity Delivered as a 50 % suspension
Required equipment
 
  • Sodium acetate trihydrate
  • Ethanol
  • Hydrochloric acid
  • Al(III)chloride, Co(II)chloride, Cu(II)chloride,Fe(III)chloride, or Zn(II)chloride
  • Nickel II sulfate
  • Sodium chloride
  • Acetic acid
  • Wash Buffer
  • Centrifuge 50 mL Tubes
  • Vortex mixer
  • End-over-end shaker

Applications - loading of the Agarose with metal ions

All and buffer compositions protocols are also avaible as PDF-Files on the Protocols & Datasheets page.
   
A.Protocol to load Nickel
 
 
  1. Transfer 20 mL PureCube NTA or IDA Agarose suspension into a 50 mL centrifuge tube.
  2. Spin the tube for 5 min at 500 x g to pellet the agarose. Remove the supernatant. Resuspend with 20 mL double distilled water.
  3. Wash two more times with 20 mL water.
  4. Wash 3x with 20 mL 50 mM sodium acetate, pH 6.0.
  5. Wash 1x with 20 mL double distilled water.
  6. Add 20 ml 2.5% nickel sulfate solution and incubate for 2 h.
  7. Wash 4x with 20 mL double distilled water.
  8. Add 20 mL Wash Buffer and incubate for 10 min.
  9. Wash 1x with 20 mL double distilled water.
  10. Wash 6x with 20 mL 20 mM Tris-HCl, pH 7.5.
  11. Wash 1x with 20 mL double distilled water.
  12. Resuspend the Ni-NTA or Ni-IDA Agarose in 20 mL Agarose Storage buffer, yielding a 50% suspension. Store at 4°C.
 B.Protocol to load Co, Cu, AL, Fe; Zn and other metals
 
 
  1. Transfer 20 mL PureCube NTA or IDA Agarose suspension into a 50 mL centrifuge tube.
  2. Spin the tube for 5 min at 500 x g to pellet the agarose. Remove the supernatant. Resuspend with 20 mL double distilled water.
  3. Wash two more times with 20 mL water.
  4. Wash 3x with 20 mL 50 mM sodium acetate, pH 6.0.
  5. Wash 1x with 20 mL double distilled water.
  6. Add 20 ml 2.5% transition metal solution and incubate for 2 h on an end-over-end shaker.
  7. Wash 4x with 20 mL double distilled water.
  8. Wash 6x with 20 mL 20 mM Tris-HCl, pH 7.5.
  9. Wash 1x with 20 mL double distilled water.
  10. Resuspend the Ni-NTA or Ni-IDA Agarose in 20 mL Agarose Storage buffer, yielding a 50% suspension. Store at 4°C.

References:

  1. Schönberger N., et al. Chromatopanning for the identification of gallium binding peptides. Elsevier; Jornal of Chromatography A, Published August 30, 2019
Agarose purification
The His tag is the most widely used affinity tag due to its small size and versatility under native and denaturing conditions, as well as in presence of detergents and many other additives. Cube Biotech offers high-performance PureCube NTA Agarose, based on BioWorks Workbeads. NTA Agarose can be loaded with transition metals to obtain different specificities for his-tagged proteins, or for other applications such as purification of phosphorylated proteins. NTA Agarose is also available as pre-packed cartridge for FPLC applications. For purification from diluted solutions, NTA MagBeads are recommended. Do you prefer IDA? We offer the same high-quality resin with that chelating ligand! (what's the difference?)

Why PureCube NTA Agarose?

PureCube NTA Agarose from Cube Biotech was successfully used in the following publications:

 Coupled molecule / metal ionPurified substanceYearAuthor
Gallium Phages 2019 Schönberger N., Braun R., Matys S., Lederer F-L., Lehmann F., Flemming K., Pollmann K.1

Features

Usage Unloaded NTA Agarose for costumized purpose
Specifity Depending of loaded metal ion (E.g. Nickel for His-affinity)
Binding capacity Depending on loaded metal Ion
Stability Stable in pH2-14; 100% methanol, 100% Ethanol, 8M urea, 6M Guanidinium hydrochloride, 30% acetonitrile
Particle diameter ~40 µm
Filling quantity Delivered as a 50 % suspension
Required equipment
 
  • Sodium acetate trihydrate
  • Ethanol
  • Hydrochloric acid
  • Al(III)chloride, Co(II)chloride, Cu(II)chloride,Fe(III)chloride, or Zn(II)chloride
  • Nickel II sulfate
  • Sodium chloride
  • Acetic acid
  • Wash Buffer
  • Centrifuge 50 mL Tubes
  • Vortex mixer
  • End-over-end shaker

Applications - loading of the Agarose with metal ions

All and buffer compositions protocols are also avaible as PDF-Files on the Protocols & Datasheets page.
   
A.Protocol to load Nickel
 
 
  1. Transfer 20 mL PureCube NTA or IDA Agarose suspension into a 50 mL centrifuge tube.
  2. Spin the tube for 5 min at 500 x g to pellet the agarose. Remove the supernatant. Resuspend with 20 mL double distilled water.
  3. Wash two more times with 20 mL water.
  4. Wash 3x with 20 mL 50 mM sodium acetate, pH 6.0.
  5. Wash 1x with 20 mL double distilled water.
  6. Add 20 ml 2.5% nickel sulfate solution and incubate for 2 h.
  7. Wash 4x with 20 mL double distilled water.
  8. Add 20 mL Wash Buffer and incubate for 10 min.
  9. Wash 1x with 20 mL double distilled water.
  10. Wash 6x with 20 mL 20 mM Tris-HCl, pH 7.5.
  11. Wash 1x with 20 mL double distilled water.
  12. Resuspend the Ni-NTA or Ni-IDA Agarose in 20 mL Agarose Storage buffer, yielding a 50% suspension. Store at 4°C.
 B.Protocol to load Co, Cu, AL, Fe; Zn and other metals
 
 
  1. Transfer 20 mL PureCube NTA or IDA Agarose suspension into a 50 mL centrifuge tube.
  2. Spin the tube for 5 min at 500 x g to pellet the agarose. Remove the supernatant. Resuspend with 20 mL double distilled water.
  3. Wash two more times with 20 mL water.
  4. Wash 3x with 20 mL 50 mM sodium acetate, pH 6.0.
  5. Wash 1x with 20 mL double distilled water.
  6. Add 20 ml 2.5% transition metal solution and incubate for 2 h on an end-over-end shaker.
  7. Wash 4x with 20 mL double distilled water.
  8. Wash 6x with 20 mL 20 mM Tris-HCl, pH 7.5.
  9. Wash 1x with 20 mL double distilled water.
  10. Resuspend the Ni-NTA or Ni-IDA Agarose in 20 mL Agarose Storage buffer, yielding a 50% suspension. Store at 4°C.

References:

  1. Schönberger N., et al. Chromatopanning for the identification of gallium binding peptides. Elsevier; Jornal of Chromatography A, Published August 30, 2019
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PureCube NTA Agarose PureCube NTA Agarose
Article number: 31701
NTA Agarose
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PureCube NTA Agarose PureCube NTA Agarose
2 ml 50% NTA Agarose - for loading with transition metals
Article number: 31701
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