FastSepô IDA Agarose for His-tagged Recombinant Proteins  (C.)
FastSepô IDA Agarose for His-tagged Recombinant Proteins (C.)

Ni2+, Co2+, Cu2+, or Zn2+ charged nitrilotriacetic acid (NTA) coupled to Agarose CL-6B via a stable and uncharged long ether hydrophilic spacer arm, and offers high binding capacity and minimal non-specific binding.

Also Available FastSep™ High Performance IDA Resins
 
Name Cat# Description Price Qty
Ni-IDA Cartridge
NIIDA-CC1X5
5 x 1ml nickel-charged IDA chromatography cartridge
$99.00
Ni-IDA Cartridge
NIIDA-CC5
1 x 5ml nickel-charged IDA chromatography cartridge
$89.00
Ni-IDA Cartridge
NIIDA-CC5X5
5 x 5ml nickel-charged IDA chromatography cartridge
$369.00
Co-IDA Cartridge
COIDA-CC1X5
5 x 1ml cobalt-charged IDA chromatography cartridge
$99.00
Co-IDA Cartridge
COIDA-CC5
1 x 5ml cobalt-charged IDA chromatography cartridge
$89.00
Co-IDA Cartridge
COIDA-CC5X5
5 x 5ml cobalt-charged IDA chromatography cartridge
$369.00
Cu-IDA Cartridge
CUIDA-CC1X5
5 x 1ml copper-charged IDA chromatography cartridge
$99.00
Cu-IDA Cartridge
CUIDA-CC5
1 x 5ml copper-charged IDA chromatography cartridge
$89.00
Cu-IDA Cartridge
CUIDA-CC5X5
5 x 5ml copper-charged IDA chromatography cartridge
$369.00
Zn-IDA Cartridge
ZNIDA-CC1X5
5 x 1ml zinc-charged IDA chromatography cartridge
$99.00
Zn-IDA Cartridge
ZNIDA-CC5
1 x 5ml zinc-charged IDA chromatography cartridge
$89.00
Zn-IDA Cartridge
ZNIDA-CC5X5
5 x 5ml zinc-charged IDA chromatography cartridge
$369.00

Description

FastSep high performance resins and chromatography cartridges

- Numerous ligand-metal ion combinations 
- High affinity & high capacity 
- Minimal non-specific binding & excellent purity 
- Long spacer arms & minimal steric hindrance 
- Low metal ion leaching & excellent stability 
- Maintain high performance after multiple repeated use 

Application

- For His-tagged protein purification 
- Simply replace your existing Ni-NTA resin or cartridge, no optimization or protocol changes necessary 
- Purification under native and denaturing conditions 
- Suitable for small proteins, large protein complexes, and proteins with low expression rates 

Purification of His-tagged Recombinant Proteins

A his-tag, or polyhistidine tag, is a string of histidine residues at either the N or C terminus of a recombinant protein. Such a string can have 4 to 10 residues, with 6 being the most common a hexahistidine tag. Recombinant proteins can have more than one hexahistidine tag. 

In IMAC, metal ions are immobilized on a resin matrix using a chelating agent such as NTA or IDA. Ni2+, Co2+, Cu2+, and Zn2+ are the most commonly used ions for his-tag purification of a recombinant protein. The his-tag has a high affinity for these metal ions and binds strongly to IMAC column, while most other proteins in the lysate will not bind to resin, or bind only weakly. The use of a his-tag and IMAC can therefore provide relatively pure recombinant protein directly from a crude lysate. 

How to choose different ligands and metal ions in IMAC?

NTA and IDA are the two most widely used ligands in IMAC. NTA is a tetravalent ligand with coordination number 4 and therefore stronger coordination of metal ions, while IDA is a trivalent ligand with coordination number 3. Generally, NTA gives higher binding specificity and lower metal ion leaching, while IDA shows more non-specific binding and increased metal ion leaching. IDA can have higher metal ion loading density and usually requires much lower imidazole concentration in eluent. 

For metal ions going from Co2+, Zn2+, Ni2+, to Cu2+, generally binding affinity increases while binding specificity decreases. Cu2+ has the highest affinity giving the highest protein recovery, but low specificity. Co2+ has the lowest affinity but highest specificity, reducing the amount of impurity proteins in eluate. Ni2+ is often most widely used because it provides a good balance between affinity and specificity. 

As many factors can affect the choice of optimal ligand-metal ion combination, the best approach is to try different combinations and let your proteins decide. FastSep(TM) high performance resins and cartridges offer numerous ligand-metal ion combinations to meet various purification needs. 
FastSep(TM) high performance resins and chromatography cartridges are based on highly cross-linked agarose 6% beads which have structural characteristics that result in superb physical strength, excellent flow properties, low backpressure, and an open pore structure. FastSep(TM) high performance resins and chromatography cartridges allow operation at very high flow rates, resulting in increased throughput and rapid column cleaning between chromatography runs. 

Specifications

Chromatography technique Histidine-tagged protein purification
Matrix  Highly cross-linked 6% beaded agarose
Active group Ni2+, Co2+, Cu2+, or Zn2+ charged Iminodiacetic Acid (IDA)
Active group density >40 umol/ml drained medium 
Binding capacity >40 mg/ml drained medium
Spacer  12 atom stable and uncharged ether hydrophilic linkage
Bead geometry & size Spherical 50 to 150 um
Bead mean diameter d50v 90 um 
Linear flow velocity <75 cm/h at 25 degC, HR 16/10 column, 5 cm bed height 
Recommended linear flow rate <30 cm/h
Pressure/flow specification Base matrix 100-200 cm/h, pressure drop cm H2O/bed height = 15, bed height 10 cm, 5 cm i.d. 
1ml Cartridge bed dimensions 7×25 mm
1ml Cartridge bed height 25 mm 
1ml Cartridge bed volume 1 ml 
1ml Cartridge column I.D 7 mm
5ml Cartridge bed dimensions 16×25 mm
5ml Cartridge bed height   25 mm
5ml Cartridge bed volume  5 ml 
5ml Cartridge column I.D 16 mm
Maximum cartridge flow rate 5 ml/min (1 ml cartridge) or 20 ml/min (5ml cartridge) 
Recommended cartridge flow rate   1 ml/min (1 ml cartridge) or 5 ml/min (5 ml cartridge)
Maximum pressure during operation 5 bar [0.5 MPa] (70 psi)
pH Stability working range 3 to 13 
pH Stability cleaning in place (cip) 2 to 14
Chemical stability Stable to commonly used aqueous solutions. Can be used with non-ionic detergents, denaturing solvents, e.g. 8 M urea and 6 M guanidine hydrochloride, Stable in organic solvents, e.g. 50% methylformamide and 50% dioxane.
Storage 2 to 8 degC
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This item: FastSepô IDA Agarose for His-tagged Recombinant Proteins (C.) [1 item(s)]
Price for all: $100.00
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