Affi-Gel Blue Gel

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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.

Structure of Affi-Gel Blue Gel
Structure of Affli-Gel blue gel
Purification Applications of Blood Proteins on Affi-Gel Blue Gel

Affi-Gel blue gel has been used to separate and purify a number of different serum and plasma proteins:

  • Purification of human serum complement
  • Purification of rat alpha fetoprotein
  • Purification of rat serum albumin
  • Separation and purification of plasma proteins
  • Purification of alpha-2 macroglobulin
  • Characterization of fragments of human albumin
  • Albumin removal from serum
  • Isolation of sarcoplasmic reticulum vesicles
  • Purification of equine thyromedin
  • Affinity separation of human plasma gelsolin
To read more click here
Affi-Gel Blue Gel Enzyme Applications
Enzyme Source Eluant
3',5' cyclic AMP phosphodiesterase Dictyostelium discoideum 0.18 M NaCl
3'-PGA diesterase Yeast 0.6 M KCl in 50 mM Tris-HCl
5-methyl-L-tetrahydrofolate reductase Bovine liver 0.4–3.0 M KCl
Acylation stimulating protein Human plasma 3 M NaCl, 0.02 M phosphate
Adenylate cyclase Bovine brain 1 mM KCl, 8 mM ATP,
16 mM MgCl2, 1 mM EDTA
Alkaline phosphatase E. coli
Alkyl hydroperoxide reductase S. typhimurium 1.0 M KCl, 0.5 M NaCl
Aspergillins (RIPs) Aspergillus 0.5–1 M NaCl gradient
ATP:AMP phosphotransferase Bovine heart 0.4–2.0 M NaCl
Calmodulin-dependent cyclic
nucleotide phosphodiesterase
Bovine brain 0.15–1.5 M NaCl
Candida acid proteinase (CAP) C. albicans 1.5 M NaCl in citrate buffer
Carbamyl phosphate synthetase Frog liver 1 mM dithiothreitol
Casein kinase I Bovine sperm 0.2–2.5 M NaCl gradient
DNA helicase S. cerevisiae 1 M NaCl
DNA polymerase Calf thymus 0–0.5 M KCl
Formamidopyrimidine-DNA glycosylase E. coli 0.1–0.8 M KCl
GAL4(63) S. cerevisiae STD, 250 mM NaCl
Glutamate dehydrogenase Yeast 10 mM NADH, 1 M NH4Cl
Glutamine synthetase S. typhimurium 5 mM ATP
Glyoxalase II Rat erythrocytes 0–0.2 M KCl
GMP reductase Human erythrocytes 1 mM NADPH, 2 mM GMP
GTP:RNA guanylytransferase Wheat germ 0.05–0.75 M NaCl
Isocitrate dehydrogenase E. coli 2 mM NADP
Isocitrate dehydrogenase E. coli 2 mM NADP+
MB creatine kinase Human heart 0.25 M NaCl
Membrane-bound phosphatidylinositol
kinase
Rat brain Triton X-100, NaCl, glycerol
Methylenetetrahydrofolate reductase Porcine liver 0–10 mM NADPH
Metmyoglobin reductase Bovine heart 1.0 M NaCl, 1 mM NADH+
Mitochondrial hexokinase Rat brain Glucose 6-phosphate
Phosphodiesterase Bovine brain 0.2 M KCl
Phosphatidic acid phosphatase Porcine thymus membranes 0.25–3 M NaCl gradient
Phosphoinositiede-specific
phospholipase C (PIC)
Bovine heart 2 M NaCl
Polyadenylate-binding protein (PABP) P. sativum 2 M Guanidine-HCl
RNA ligase E. coli 0.2 M NaCl, 2 mM ATP
Serine transhydroxymethylase Porcine liver 0.5 M KCl
Thymidylate synthetase S. cervisiae 0.25–1 M KCl
Tyrosine phenollyase Erwinia herbicola 1.5 M NaCl,
0.5 mM mercaptoethanol

To read more click here

Removal of Albumin

Comparison of Cibacron Blue Affinity-Based Formats for Depletion of Albumin From Samples Prior to Two-Dimensional Gel Electrophoresis

Test Sample Spot Number % Change % Albumin Depletion
Control 670 N/A N/A
Batch Binding 620 -6% 45%
Gel loader columns 650 -3% 36%
Aurum serum protein mini kit 700 +4% >80%

Affi-Gel blue affinity gel is a beaded, crosslinked agarose gel with covalently attached Cibacron Blue F3GA dye. It contains >1.9 mg dye per ml of gel and has a typical albumin capacity of >11 mg/ml. Affi-Gel Blue gel purifies a large range of proteins from widely divergent origins. The blue dye functions as an ionic, hydrophobic, aromatic, or sterically active binding site in various applications.

Affi-Gel Blue is ideally suited for albumin removal (using 50–100 mesh) and enzyme purification (using 100–200 mesh). Proteins and peptides are bound and released with a high degree of specificity by manipulating the composition of the eluent buffers.

For more information, request bulletin 1107.

Functional group Cibacron blue
Ligand concentration ≥ 1.9 mg/ml
Typical protein capacity > 11 mg/ml
pH stability 4–11
Pressure limit 15 psi
Flow rate 15–25 cm/hr
Bead size:
   50–100 mesh
   100–200 mesh

150–300 µm
75–150 µm
Storage 4°C
Shipping buffer 0.05 % NaN3 in water
Sanitization 0.1 M NaOH
Affi-Gel Blue Gel

153-7301
100 ml, 50–100 mesh, ready-to-use crosslinked agarose bead with covalently attached Cibacron Blue F3GA dye

List Price:   $512.00
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Affi-Gel Blue Gel

153-7302
100 ml, 100–200 mesh, ready-to-use crosslinked agarose bead with covalently attached Cibacron Blue F3GA dye

List Price:   $512.00
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Quantity:   Add to cart
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Number Description Options
1107 Affi-Gel Blue Affinity Chromatography Gel for Enzyme and Blood Protein Purifications Click to download
4110139 Instruction Manual, Aurum Affi-Gel Blue Mini Kits and Columns Sample Preparation, Rev B Click to download
LIT590 Instruction Manual, Affi-Gel Blue Gel Click to download [ Add to Cart (Free) ]
3168 Comparison of Cibacron Blue Affinity-Based Formats for Depletion of Albumin From Samples Prior to Two-Dimensional Gel Electrophoresis Click to download