Profinity IMAC Resins

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Overview

Affinity Media Selection Guide
  Matrix Functional Group Specificity Capacity Working pH Pressure Limit Applications
Ready-to-Use Affinity Media
Nuvia™ IMAC High capacity, pressure-stable polymer based on UNOsphere beads NTA charged with Ni2+ Histidine ≥40 mg/ml* 2–14 45 psi (3.1 bar) Purification of recombinant histidine-tagged proteins; can be charged with other transition metals
Profinity™ IMAC Pressure-stable polymer based on UNOsphere beads IDA, provided charged with Ni2+ and uncharged Histidine ≥15 mg/ml* 1–14 100 psi (6.8 bar) Purification of recombinant His-tagged proteins; can be charged with other transition metals
Profinity GST** Pressure-stable polymer based on UNOsphere beads Immobilized glutathione GST-tagged proteins ≥10 mg/ml 1–14 45 psi (3.1 bar) Purification of recombinant GST-tagged proteins
Profinity eXact*** Crosslinked 6% agarose Subtilisin protease Subtilisin prodomain ≥3 mg/ml tag-free protein 2–10 10 psi (minus system pressure) Generation of native, tag-free protein by on-column purification and cleavage
Affi-Gel® Protein A Crosslinked agarose Protein A
2 mg/ml
IgG   2–10 15 psi (1 bar) Purification of IgG from ascites, serum, and culture fluid; with MAPS buffer system, purification of 10 mg mouse IgG, per ml of gel is possible
Affi-Prep® Protein A Pressure-stable polymer Protein A
2 mg/ml
IgG   2–10 1,000 psi (70 bar) Purified IgG from ascites, serum, and culture fluid; pressure-stable support for process-scale applications
Affi-Gel Blue Crosslinked agarose Cibacron Blue F3GA 1.9 mg/ml Albumin; general ≥11 mg/ml 2–10 15 psi (1 bar) Binds many nucleotide-requiring enzymes, albumin, and other proteins
DEAE Affi-Gel Blue Crosslinked agarose Cibacron Blue F3GA and DEAE Albumin and serum protiens 0.14 ml serum/ml gel 2–10 15 psi (1 bar) Purifies protease-free IgG from ascites, serum, and culture fluid with minimal sample preparation
CM Affi-Gel Blue Crosslinked agarose Cibacron Blue F3GA and CM Albumin and serum protiens 0.17–0.5 ml serum/ml gel 2–11 15 psi (1 bar) Produces albumin and protease-free antibody preparation from serum without prior dialysis
Affi-Prep polymyxin Pressure-stable polymer Polymyxin 2–4 mg/ml Endotoxins >5 mg/ml 2–10 1,000 psi (70 bar) Endotoxin removal
Affi-Gel boronate Polyacrylamide gel Boronate 1.05 ± 0.15 cis-diols

meq/g
130 µmol sorbitol/ml 2–10 15 psi (1 bar) Adsorption of cis-hydroxyl-containing molecules, including sugars, nucleotides, and glycopeptides
Activated Media for Spontaneous Ligand Immobilization
Profinity epoxide Pressure-stable polymer based on UNOsphere beads Epoxy group Nucleophiles; amini, thiol, -COOH 36–40 mg/ml lgG 1–14 Up to 80 psi (5.5 bar) Activated matrix for the immobilization of barious ligands (for example, protein A, StrepTactin, and immunoglobulins)
Affi-Gel 10 Crosslinked agarose N-hydroxy-succinimide ≥10 µmol/ml -NH2 35 mg/ml 3–11 15 psi (1 bar) For coupling proteins with pI 6.5–11
Affi-Gel 15 Crosslinked agarose N-hydroxy-succinimide ≥9 µmol/ml -NH2 35 mg/ml 3–11 15 psi (1 bar) For coupling proteins with pI 6.5–11
Affi-Gel Hz Crosslinked agarose Hydrazide Oxidized carbohydrates 1–5 mg/ml 2–10 15 psi (1 bar) Immobilization of immunoglobulins and other glycoproteins via carbohydrate molecules
Affinity Media Using Carbodiimide Activation
Affi-Gel 102 Crosslinked agarose -NH2 16 ± 4 meq/ml -COOH 40 mg 2–11 15 psi (1 bar) Carbodimide coupling of carboxyl-containing ligand

* Refer to bulletin 3193 for purification conditions.
** For example, Profinity GST resin is only available in prepacked cartridges.
***Profinity eXact purification resin.

Chemical Compatibilities for Profinity IMAC Resins

Chemical compatibilities for binding of target protein to Profinity IMAC Ni-charged resin*

Reagent Concentration**
Buffer Reagents
Tris 50 mM
HEPES 50 mM
MOPS 50 mM
Sodium or potassium phosphate 50 mM
Chelating Agents
EDTA, EGTA 0.1 mM
Sulfhydryl Reagents
β-Mercaptoethanol 30 mM
DTT 5 mM
TCEP 10 mM
Detergents
Nonionic detergents
(Triton, Tween, NP-40)
5%
Cationic detergents (CTAB) 1% (care must be taken to avoid protein precipitation)
Zwitterionic detergents
(CHAPS, CHAPSO)
5%
Anionic detergents
(SDS, Sarkosyl)
1%
Denaturing Agents
Guanidine HCl 6-M
Urea 8-M
Other Additives
NaCl 2 M (at least 300 mM NaCl should be included in buffers)
MgCl2 100 mM (HEPES or Tris should be used to prevent precipitation)
CaCl2 10 mM (HEPES or Tris should be used to prevent precipitation)
Glycerol 20%
Ethanol 20%
Imidazole 5–10 mM in wash buffer
500 mM for elution
Citrate 80 mM

* In order to determine the efficacy of protein binding and recovery, tests were performed using a purified 75 kD protein in a static mode. All tests were carried out using Micro Bio-Spin columns packed with Profinity IMAC resin and eluted by centrifugation.
** Profinity IMAC binding capacities are unaffected with typical reagents used for histidine-tagged protein purification, up to the concentrations given.

Comparison of Maximum Operating Pressures

Comparison of maximum operating pressure for different IMAC resins. The pressure values shown are the estimated maximum operating pressures for each resin. Beyond these pressures, the column beds are seriously compressed and damaged. Resins were converted to 50% (v/v) slurry in 20 mM sodium phosphate buffer and packed in a 1.1 x 20 cm column to a bed height of 20 cm up to 42 psi (3 bar). Flow rates were then increased stepwise by 200 cm/hr and held for 2 min at each step. The pressure-flow curve for Profinity IMAC resin became nonlinear only at pressures above 109 psi (7.5 bar), the point defined as the intersection of the two tangents on the pressure-flow curve.

Profinity IMAC resin is an affinity chromatography support for the purification of recombinant histidine (His)-tagged proteins. Immobilized metal affinity chromatography (IMAC) is a powerful purification technique that relies on the affinity that His has for immobilized transition metals. Profinity IMAC resin, based on UNOsphere beads, contains iminodiacetic acid (IDA) as the chelating ligand for di- or trivalent metal ions. Its chemical structure allows highly selective binding of recombinant His-tagged proteins when charged with Ni2+ or other transition metals, such as Zn2+ or Cu2+.

Profinity IMAC resin offers high capacity at high flow rates. It can be used under either nondenaturing or denaturing conditions and is suitable for purification using liquid chromatographic instrumentation, gravity-flow columns, or spin columns.

The resin is available in two forms: uncharged and precharged with Ni2+. The uncharged form can be charged with the metal ion of your choice for even greater purification flexibility.

Features of Profinity IMAC resins include:

  • Optimal ligand density for higher purity of target protein
  • Superb mechanical strength
  • Excellent pressure-flow properties for high maximum operating pressures and high flow rates, which allow rapid purification as well as rapid column cleaning and re-equilibration
  • Stable from pH 1 to 14
  • Compatible with denaturing agents, detergents, and reducing agents

For more information, request bulletin 5456.

Larger volumes and special packaging for industrial applications are available upon request.

Profinity IMAC Uncharged Resin

156-0121
10 ml, ready-to-use support for immobilized metal affinity chromatography

List Price:   $81.00
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Profinity IMAC Uncharged Resin

156-0123
50 ml, ready-to-use support for immobilized metal affinity chromatography

List Price:   $344.00
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Profinity IMAC Uncharged Resin

156-0125
500 ml, ready-to-use support for immobilized metal affinity chromatography

List Price:   $3,030.00
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Profinity IMAC Uncharged Resin

156-0127
1 L, ready-to-use support for immobilized metal affinity chromatography

List Price:   $4,584.00
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Profinity IMAC Ni-Charged Resin

156-0131
10 ml, ready-to-use support for immobilized metal affinity chromatography

List Price:   $100.00
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Profinity IMAC Ni-Charged Resin

156-0133
25 ml, ready-to-use support for immobilized metal affinity chromatography

List Price:   $235.00
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Profinity IMAC Ni-Charged Resin

156-0135
100 ml, ready-to-use support for immobilized metal affinity chromatography

List Price:   $893.00
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Profinity IMAC Ni-Charged Resin

156-0137
500 ml, ready-to-use support for immobilized metal affinity chromatography

List Price:   $4,048.00
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Number Description Options
5456 Profinity IMAC Resin: An Optimal Support for His-Tagged Protein Purification, Rev A Click to download
3193 Profinity IMAC Resin Product Information Sheet Click to download
5283 Profinity IMAC Ni-Charged Resin Flier, Rev A Click to download
5251 Rapid Method Development With the BioLogic DuoFlow Chromatography System for the Purification of His-Tagged Proteins, Rev A Click to download
5501 Rapid, Efficient Purification and Evaluation of His-Tagged Proteins, Rev A Click to download
5444 Bio-Scale Mini Profinity IMAC Cartridge Product Information Sheet, Rev A Click to download
5514 Automated Purification of a His-Tagged Protein With the Profinia Protein Purification System: Comparison With Another Low-Pressure Chromatography System, Rev A Click to download [ Add to Cart (Free) ]
10001677 Instruction Manual, Profinity IMAC Resins, Rev B Click to download
10005455 Instruction Manual, Bio-Scale™ Mini Profinity™ IMAC Cartridges, 1 and 5 ml, Rev C Click to download