Affi-Gel Boronate Gel

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Affi-Gel Boronate Gel

Overview

Affinity Chromatography Resin 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, available charged with Ni2+ and uncharged Histidine ≥15 mg/ml* 1–14 100 psi
(6.8 bar)
Suitable for purification of larger and difficult-to-purify recombinant histidine-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 proteins 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 proteins 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 proteins 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 various 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)
Carbodiimide coupling of carboxyl-containing ligand

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

Boronate Applications
Application Application Buffer Molecules in Vo Retained Molecules Elution Buffer Support Used
Adenylate cyclase assay HEPES pH 7.5 + MgCl2 cAMP ATP, AMP, and adenosine 0.05 M NaOAc Bio-Gel P-150 boronate gel
Isolation of catecholamines from urine 0.1 M phosphate pH 7.0 + EDTA Other urine components and DOPA Norepinephrine, epinephrine 0.025 N HCI Boric acid gel (Sigma-Aldrich)
Modified nucleosides in urine 0.25 M NH4OAc, pH 8.8 Thymidine, adenine Pseudouridine 0.1 M HCOOH Bio-Gel P-2 gel, 200–400 mesh, boronate
Separation of mono- and oligonucleotides Triethyl ammonium Deoxyribonucleotides Ribonucleotides H2O and others Dihydroxyboryl methacrylate
Sugars 0.05 M N-methyl-morpholinium-Cl, pH 7.5 + 1 M NaOAc Erythritol, adonitol, sucrose, and D-glucose L-arabinitol, xylitol, D-mannitol, dulcitol, sorbitol, and D-fructose* Isocratic run, elution buffer same as application buffer Dihydroxyboryl cellulose
Assay of ribonucleotide reductase Tris, MgCl2 dUMP, dCMP CDP Citrate Dihydroxyboryl Cellulose Dowex 1(AG) resin

* In order of increasing retention by support.

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Structure

Affi-Gel boronate-derivatized polyacrylamide gel has affinity for coplanar adjacent cis-hydroxyl groups (cis-diols) and a high binding capacity, which provides highly efficient separation of low molecular weight molecules, such as nucleotides, nucleosides, catecholamines, and sugars. It has a sorbitol capacity of 130 µmol/ml.

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Affi-Gel® boronate-derivatized polyacrylamide gel has affinity for coplanar adjacent cis-hydroxyl groups (cis-diols) and a high binding capacity, which provides highly efficient separation of low molecular weight molecules such as nucleotides, nucleosides, catecholamines, and sugars. It has a sorbitol capacity of 130 µmol/ml.

Key Features

  • High boronate capacity — 1.05 ± 0.15 meq/g
  • High diol capacity — binds 130 µmol sorbitol/ml gel
  • Spherical polyacrylamide matrix — excellent flow rate, resolution, and reproducibility
  • Low nonspecific absorption, optimum porosity — low exclusion limit precludes significant binding of high molecular weight molecules while allowing rapid equilibration and binding of low and medium molecular weight molecules

Preparation and Use

The gel is supplied as dried material and must be hydrated in an appropriate buffer. Buffer choice is dependent upon application. Buffers such as Tris should be avoided as they can adversely affect binding capacity (Schott et al.1973). The presence of Mg2+ may enhance binding. In general, binding is effected at pH > 7.5 and elution at pH < 6.5. boric acid, sorbitol, or mannitol can also be used for elution.

For more information, request bulletin 1066, which can be found under the Documents tab.

 

 

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Affi-Gel Boronate Gel

1536103
5 g, ready-to-use affinity chromatography support, for small MW molecules (nucleotides, nucleosides, catecholamines, and sugars)

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Affi-Gel Boronate Gel

153-6104
50 g, boronate affinity chromatography media, for small MW molecules (nucleotides, nucleosides, catecholamines, and sugars)

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1066 Affi-Gel® Boronate Affinity Gel to Separate Ribonucleotides, Ribonucleosides, Sugars, Catecholamines, and Coenzymes, Rev B Click to download

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