Affi-Gel Blue Gel

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Overview

Structure of Affi-Gel Blue Gel
Structure of Affi-Gel Blue Gel
Structure of Affli-Gel blue gel
Applications
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
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Enzyme Applications
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

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Removal of Albumin
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:   $500.00
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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:   $500.00
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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 ]
3168 Comparison of Cibacron Blue Affinity-Based Formats for Depletion of Albumin From Samples Prior to Two-Dimensional Gel Electrophoresis Click to download