Purification Media, BioSeparation™ Affinity, Heparin
|BioSeparation?||Storage: 4°C Do not freezeShipping: RT|
Heparin Beads are made by coupling 20-35kD size Heparin extracted from Porcine intestinal mucosa (Type I) to Sepharose beads. These beads are useful for isolating heparin-binding protein such as Antithrombin III, lipoproteins, various enzymes and DNA binding proteins.
Heparin Beads Specifications:
Matrix: Sepharose CL-4B
Coupling method: TargetLock.
Heparin density: 0.7-1.2mg/ml of 20-35kD Heparin. Thrombin binding capacity minimum 0.7mg/ml of beads
Mean bead size: 40 -165um
Range of pH: 3.0-11.0
Bead structure: Highly cross-linked spherical agarose, 4%
Max back pressure: 0.3MPa, 3 bar
Max. flow rates: 4ml/min/cm2
Recommended flow rate: 1-3ml/min/cm2
Storage and Stability:
May be stored at 4°C. For long-term storage, aliquot and store at 4°C. Do not freeze. Aliquots are stable for 6 months. For maximum recovery of product, centrifuge the original vial prior to removing the cap. Further dilutions can be made in assay buffer.
Protocol: Salt purification of Heparin-binding proteins:
1. Invert bottle several times to mix beads in storage buffer.
2. Pack column with 2-4ml beads
3. Equilibrate column with 10mM Tris-HCl pH 7.5. Matrix is robust -You may modify buffer to include reducing agents, urea, guanidine, sodium thiocyanate and detergents as needed.
4. Load you protein sample on column at 0.5-1.0ml/min.
5. Wash column with approximately 30 volumes of the equilibration buffer.
6. Elute bound proteins by applying high salt buffer (10mM Tris-HCl pH 7.5 added with sodium chloride to 1.5N or other commonly used salts such as KCl and Ammonium sulfate).
7. Wash column with 20 volumes of ddH2O to regenerate it.
8. Equilibrate column with 10mM Tris-HCl pH 7.5.
9. Store column at 4°C in PBS pH 7.4, 0.3N sodium chloride added with 0.01% Thermasol as a preservative.
Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.