IDA Agarose


Purification resins
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 IDA Agarose, based on BioWorks Workbeads. IDA 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. IDA Agarose is also available as pre-packed cartridge for FPLC applications. For purification from diluted solutions, IDA MagBeads are recommended. For higher specificity, we offer the same high-quality resin with an IDA ligand. (what's the difference?)

Why PureCube IDA Agarose?

  • Can be loaded with most transition metals
  • Stable under commonly used concentrations of DTT and EDTA
  • Can be regenerated for reuse

Features

Usage Unloaded IDA 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 protocols and buffer compositions are also avaible as PDF-Files on the Protocols & Datasheets page.
   
A.Protocol to load Nickel
 
 
  1. Transfer 20 mL PureCube 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-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 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-IDA Agarose in 20 mL Agarose Storage buffer, yielding a 50% suspension. Store at 4°C.
Purification resins
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 IDA Agarose, based on BioWorks Workbeads. IDA 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. IDA Agarose is also available as pre-packed cartridge for FPLC applications. For purification from diluted solutions, IDA MagBeads are recommended. For higher specificity, we offer the same high-quality resin with an IDA ligand. (what's the difference?)

Why PureCube IDA Agarose?

  • Can be loaded with most transition metals
  • Stable under commonly used concentrations of DTT and EDTA
  • Can be regenerated for reuse

Features

Usage Unloaded IDA 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 protocols and buffer compositions are also avaible as PDF-Files on the Protocols & Datasheets page.
   
A.Protocol to load Nickel
 
 
  1. Transfer 20 mL PureCube 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-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 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-IDA Agarose in 20 mL Agarose Storage buffer, yielding a 50% suspension. Store at 4°C.
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PureCube IDA Agarose PureCube IDA Agarose
Article number: 30703
IDA Agarose
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PureCube IDA Agarose PureCube IDA Agarose
2 ml 50% IDA Agarose - for loading with transition metals
Article number: 30703
Sales price: From €68.00 *
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