Affinity Chromatography Analysis

AFpak ACA-894 (an affinity column) has as the ligand Concanavalin A (a lectin type). This column is recommended for the analysis of polysaccharides and glycoproteins.

Sample : Ovalbumin

Column       : Shodex AFpak ACA-894 (8.0mmID*50mm)
Eluent       : (A); 0.02M Tris-HCl(pH7.4) + 0.5M NaCl
               (B); (A) + 0.05M alpha-Methyl-D(+)-glucoside
               Linear gradient: (A) to (B) for 60min
Flow rate    : 0.3mL/min
Detector     : UV(280nm)
Column temp. : Room temp.

AFpak ADS-894 (an affinity column) with a dextran sulfate ligand is recommended for the analysis of lipoproteins and coagulant components in blood.

Sample : alpha1 Antitrypsin

Column       : Shodex AFpak ADS-894 (8.0mmID*50mm)
Eluent       : (A); 0.02M Tris-HCl(pH7.4)
               (B); (A) + 1.0M NaCl
               Linear gradient: (A) to (B) for 60min 
Flow rate    : 1.0mL/min
Detector     : UV(280nm)
Column temp. : Room temp.

AFpak ACB-894 (an affinity column) with a cibacron blue ligand is recommended for the analysis of albumin and NAD-dependent enzymes.

Sample : Bovine serum albumin

Column       : Shodex AFpak ACB-894 (8.0mmID*50mm)
Eluent       : (A); 0.1M Potassium phosphate buffer(pH5.0)
               (B); 0.1M Potassium phosphate buffer(pH7.5) + 1.5M KCl
               Step gradient: (A) to (B)
Flow rate    : 1.0mL/min
Detector     : UV(280nm)
Column temp. : Room temp.

AFpak AHR-894 (an affinity column) with a heparin ligand is recommended for the analysis of lipoproteins and coagulant components in blood.

Sample :
1. Ovalbumin
2. Conalbumin
3. Lysozyme

Column       : Shodex AFpak AHR-894 (8.0mmID*50mm)
Eluent       : (A); 0.01M Tris-HCl(pH7.5) + 0.01M NaCl
               (B); (A) + 0.3M NaCl
               Linear gradient: (A) to (B)
Flow rate    : 0.5mL/min
Detector     : UV(280nm)
Column temp. : 25deg-C

AFpak AAV-894 an affinity column has avidin (a protein in egg white) as a ligand. This column is recommended for the analysis of biotin derivatives.

Sample : Biotin 4-aminobenzoic acid

Column       : Shodex AFpak AAV-894 (8.0mmID*50mm)
Eluent       : (A); 0.01M Sodium acetate buffer(pH7.0) + 0.15M NaCl
               (B); 0.1M Acetic acid/Isopropanol=50/50
               Step gradient: (A) to (B)
Flow rate    : 1.0mL/min
Detector     : UV(260nm)
Column temp. : Room temp.

AFpak AGT-894 (an affinity column) has glutathione (sulfohydrate present in large quantity in living organisms) as a ligand.

Sample : Glutathione S-transferase

Column       : Shodex AFpak AGT-894 (8.0mmID*50mm)
Eluent       : (A); 22mM Phosphate buffer(pH7.0)
               (B); 50mM Tris buffer(pH9.6) + 5mM Glutathione
               Step gradient: (A) to (B)
Flow rate    : 1.0mL/min
Detector     : UV(280nm)
Column temp. : Room temp.

L-Lactate dehydrogenase was analyzed using AFpak ANA-894 (an affinity column).

Sample : L-Lactate dehydrogenase

Column       : Shodex AFpak ANA-894 (8.0mmID*50mm)
Eluent       : (A); 0.01M Potassium phosphate buffer(pH7.0)
               (B); (A) + 0.5M KCl
               Step gradient: (A) to (B)
Flow rate    : 1.0mL/min
Detector     : UV(280nm)
Column temp. : Room temp.

Super oxide dismutase (SOD) was analyzed using AFpak AIA-894 (an affinity column).

Sample : Super oxide dismutase

Column       : Shodex AFpak AIA-894 (8.0mmID*50mm)
Eluent       : (A); 0.1M Sodium acetate buffer(pH7.7) + 0.5M NaCl 
               (B); 0.05M Tris-HCl(pH8.0) + 0.15M NH4Cl 
               Step gradient: (A) to (B)
Flow rate    : 0.5mL/min
Detector     : UV(280nm)
Column temp. : Room temp.

Fibrinogen was analyzed using AFpak ALC-894 (an affinity column).

Sample : Fibrinogen (Sigma)

Column       : Shodex AFpak ALC-894 (8.0mmID*50mm)
Eluent       : (A); 0.05M Tris-HCl(pH7.4) + 0.15M NaCl + 0.5mM CaCl2 + 0.5mM MnCl2 
               (B); (A) + 0.1M Methyl mannoside
               Step gradient: (A) to (B)
Flow rate    : 0.5mL/min
Detector     : UV(280nm)
Column temp. : Room temp.

Plasminogen was analyzed using AFpak ALS-894 (an affinity column).

Sample : Plasminogen (from bovine plasma, Sigma)

Column       : Shodex AFpak ALS-894 (8.0mmID*50mm)
Eluent       : (A); 0.1M Sodium acetate(pH7.4)
               (B); 0.1M Sodium acetate + 0.2M 6-aminohexanoic acid(pH4.0)
               Step gradient: (A) to (B)
Flow rate    : 1.0mL/min
Detector     : UV(280nm)
Column temp. : Room temp.

Human Serum was analyzed using AFpak APG-894 (an affinity column).

Sample : Human serum

Column       : Shodex AFpak APG-894 (8.0mmID*50mm)
Eluent       : (A); 0.2M Sodium phosphate buffer(pH4.0)
               (B); 0.1M Sodium citrate buffer(pH2.5)
               Step gradient: (A) to (B)
Flow rate    : 1.0mL/min
Detector     : UV(280nm)
Column temp. : Room temp.

Bovine gamma-globulin were analyzed using AFpak APR-894 (an affinity column).

Sample :
Bovine gamma-Globulin (Sigma)

Column       : Shodex AFpak APR-894 (8.0mmID*50mm)
Eluent       : (A); 0.01M Sodium phosphate buffer(pH6.5)
               (B); (A) + 1.0M NaCl
Flow rate    : 1.0mL/min
Detector     : UV(280nm)
Column temp. : Room temp.

AFpak APA-894 (an affinity column) which has protein A as the ligand was used for the adsorption and elution of bovine gamma-globulin.

Sample : Bovine gamma-Globulin

Column       : Shodex AFpak APA-894 (8.0mmID*50mm)
Eluent       : (A); 0.1M Sodium phosphate buffer(pH4.5)
               (B); 0.1M Sodium phosphate buffer(pH3.0)
Flow rate    : 1.0mL/min
Detector     : UV(280nm)
Column temp. : Room temp.

Human gamma-globulin was analyzed using AFpak APR-894 (an affinity column).

Sample : Human gamma-Globulin (Sigma)

Column       : Shodex AFpak APR-894 (8.0mmID*50mm)
Eluent       : (A); 0.01M Sodium phosphate buffer(pH6.5)
               (B); (A) + 1.0M NaCl
               Linear gradient: (A) to (B)
Flow rate    : 1.0mL/min
Detector     : UV(280nm)
Column temp. : Room temp.

Human IgM was analyzed using AFpak APR-894 (an affinity column).

Sample : Human IgM (Sigma)

Column       : Shodex AFpak APR-894 (8.0mmID*50mm)
Eluent       : (A); 0.02M Sodium phosphate buffer(pH6.5)
               (B); (A) + 1.0M NaCl
               Linear gradient: (A) to (B)
Flow rate    : 1.0mL/min
Detector     : UV(280nm)
Column temp. : Room temp.

Human serum albumin was analyzed using AFpak APD-894 (an affinity column).

Sample : Human serum albumin (Sigma)

Column       : Shodex AFpak APD-894 (8.0mmID*50mm)
Eluent       : (A); 0.02M Tris-HCl buffer(pH5.0)
               (B); (A) + 2.0M KCl
               Step gradient: (A) to (B) 
Flow rate    : 1.0mL/min
Detector     : UV(280nm)
Column temp. : Room temp.

C-Reactive protein was analyzed using AFpak APE-894 (an affinity column).

Sample : C-Reactive protein (Sigma)

Column       : Shodex AFpak APE-894 (8.0mmID*50mm)
Eluent       : (A); Tris-HCl(pH7.0) + 0.15M NaCl + 1mM CaCl2 
               (B); (A) + 0.2M Phosphorylcholine
               Step gradient: (A) to (B) 
Flow rate    : 1.0mL/min
Detector     : UV(280nm)
Column temp. : Room temp.

The ligand of AFpak AAB-894 (an affinity column) is aminobenzamidine (serine protease inhitor). The column was used for the analysis of trypsin. Trypsin was adsorbed by the ligand when the eluent of pH 7.0 was flowed and it was released from the ligand when the eluent of pH 2.8 was flowed.

Sample : Trypsin

Column       : Shodex AFpak AAB-894 (8.0mmID*50mm)
Eluent       : (A); 0.1M Sodium acetate buffer(pH 7.0) + 0.5M NaCl
               (B); 0.1M Sodium acetate buffer(pH 2.8) + 0.5M NaCl
               Step gradient: (A) to (B)   
Flow rate    : 1.0mL/min
Detector     : UV(280nm)
Column temp. : Room temp.

Adenosin-5′-monophosphate, adenosin-5′-diphosphate and adenosin-5′-triphosphate were separated using AFpak AAF-894 (an affinity column) with 0.1M Ethylmorpholine-acetate (pH7.0) as the eluent.

Sample : Adenosin phosphate
1. Adenosin-5′-monophosphate, AMP
2. Adenosin-5′-diphosphate, ADP
3. Adenosine-5′-triphosphate, ATP

Column       : Shodex AFpak AAF-894 (8.0mmID*50mm)
Eluent       : 0.1M Ethylmorpholine-acetate buffer(pH7.0)
Flow rate    : 1.0mL/min
Detector     : UV(260nm)
Column temp. : Room temp.

Alubumin works as a trnsport protein in blood and can distinguish optical isomers. The ligand of AFpak ABA-894 (an affinity column) is BSA(bovine serum albumin). The column was used to separate optical isomers(D and L) of N-(3,5dinitrobenzoyl)-DL-phenyl glycine.

Sample : N-(3,5dinitrobenzoyl)-DL-phenyl glycine

Column       : Shodex AFpak ABA-894 (8.0mmID*50mm)
Eluent       : 0.02M Tris-HCl buffer(pH 7.0) + 0.1M NaCl 
Flow rate    : 0.5mL/min
Detector     : UV(254nm)
Column temp. : Room temp.

The ligand of AFpak ABT-894 (an affinity column) is biotion derivative of which affinity with avidin is weakened. The column was used for the analysis of avidin. Avidin is a glycoprotein and has strong affinity with biotin and can be used for the quantitation of biotin. In order to release avidin which was adsorbed by biotin, the gradient to 1M NaCl is neccessary.

Sample : 0.1% Avidin, 10micro-L

Column       : Shodex AFpak ABT-894 (8.0mmID*50mm)
Eluent       : (A); 0.01M Sodium phosphate buffer(pH 7.0)
               (B); (A) + 1.0M NaCl
               Linear gradient: 0min to 20min, (A), 20min to 40min, (A) to (B), after 40min, (B) 
Flow rate    : 0.5mL/min
Detector     : UV(230nm)
Column temp. : Room temp.

The ligand of AFpak AST-894 (an affinity column) is soybean trypsin inhibitor (STI). The column was used to adsorb and elute chymotrypsin which is a typical digestive enzyme.

Sample : Chymotrypsin

Column       : Shodex AFpak AST-894 (8.0mmID*50mm)
Eluent       : (A); 0.1M Sodium acetate buffer(pH7.7) + 0.5M NaCl
               (B); 0.1M Sodium acetate buffer(pH2.8) + 0.5M NaCl
               Linear gradient:  0min to 20min, (A), 20min to 40min, (A) to (B), after 40min, (B)
Flow rate    : 1.0mL/min
Detector     : UV(280nm)
Column temp. : Room temp.

AFpak AGA-894 ( an affinity column ) which has N-acetyl-glucosamine as the ligand was used for the adsorption and elution of lysozyme.

Sample : Lysozyme

Column       : Shodex AFpak AGA-894 (8.0mmID*50mm)
Eluent       : (A): 0.01M TRis-HCl buffer(pH7.5) + 0.01M NaCl
               (B): 0.01M TRis-HCl buffer(pH7.5) + 0.5M NaCl 
               Step gradient: 0min to 12min, (A) to (B)
Flow rate    : 0.5mL/min
Detector     : UV(280nm)
Column temp. : Room temp.

AFpak AHR-894 (an affinity column) has heparin as the ligand and is recommended for the analysis of lipoproteins and coagulant components in blood.

Sample : Lysozyme

Column       : Shodex AFpak AHR-894 (8.0mmID*50mm)
Eluent       : (A); 0.01M Tris-HCl buffer(pH7.5) + 0.01M NaCl
               (B); 0.01M Tris-HCl buffer(pH7.5) + 0.3M NaCl
               Step gradient: (A) to (B)
Flow rate    : 1.0mL/min
Detector     : UV(280nm)
Column temp. : Room temp.

AFpak AGE-894 (an affinity column) which has gelatin as the ligand was used to adsorb and elute fibronectin. Fibronectin is a glycoprotein which works to adhere and/or enlarge cell.

Sample : Fibronectin

Column       : Shodex AFpak AGE-894 (8.0mmID*50mm)
Eluent       : (A); 50mM Tris-HCl buffer(pH7.4) + 0.01M NaCl
               (B); (A) + 4M urea
               Step gradient: (A) to (B)
Flow rate    : 0.3mL/min
Detector     : UV(280nm)
Column temp. : Room temp.

Ovomucoid is a glycoprotein exists in egg white and is a trypsin inhibitor. AFpak AOV-894 which has ovomucoid as the ligand was used for the adsorption and elution of trypsin.

Sample :Trypsin (type I)

Column       : Shodex AFpak AOV-894 (8.0mmID*50mm)
Eluent       : (A); 0.1M Sodium acetate buffer(pH7.7) + 0.5M NaCl
               (B); 0.1M Sodium acetate buffer(pH2.8) + 0.5M NaCl
               Step gradient: (A) to (B)
Flow rate    : 1.0mL/min
Detector     : UV(280nm)
Column temp. : Room temp
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