Heparin Agarose 6FF
Based on 1 Customer Validation
MCE Heparin Agarose 6FF is suitable for the separation and purification of heparin-binding biomolecules, including antithrombin III, coagulation factors, other plasma proteins, DNA-binding proteins, lipoproteins, protein synthesis factors, nucleic acid-related enzymes, and steroid receptors.
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Storage :
4°C, 2 years
Do not dry or freeze
- Manual (3644KB)
- COA
To place orders, for customer services and technical support, please contact: MedChemExpress USA
Tel: 609-228-6898 E-mail: [email protected] [email protected]
Description & Advantages
Heparin is a naturally occurring glycosaminoglycan composed of alternating residues of hexuronic acid (either D-glucuronic acid or L-iduronic acid) and D-glucosamine. This polymer is highly sulfated, with N-sulfate groups attached at the C-2 position of the glucosamine units, along with various O-sulfate groups at different positions.
MCE Heparin Agarose 6FF is based on high-flow agarose microspheres, with heparin covalently coupled to the agarose matrix via an epoxy activation process. It offers excellent chemical stability, minimal ligand leakage, long service life, and ease of use.
This resin has broad applications and is suitable for the separation and purification of heparin-binding biomolecules, including antithrombin III, coagulation factors, other plasma proteins, DNA-binding proteins, lipoproteins, protein synthesis factors, nucleic acid-related enzymes, and steroid receptors.
Each 1 mL of total volume contains 0.5 mL of resin. Thoroughly resuspend the resin before use and mix well before aliquoting.
The specifications of the product correspond to the actual resin volume, with the resin content of 50%.
Protocol
Recommended Buffers (Not Provided)
| Buffer | Composition |
|---|---|
| Equilibration/Wash Buffer | 20 mM Tris-HCl, pH 7.4 |
| Elution Buffer | 20 mM Tris-HCl containing 0-2 M NaCl, pH 7.4 |
| Storage Buffer | 1× PBS containing 20% ethanol |
Note: Prepare buffers with high-purity water and filter them through a 0.22 μm or 0.45 μm membrane. Optimize the pH, salt concentration, and elution conditions according to the target molecule.
Sample Preparation
Clarify and filter samples containing heparin-binding proteins or other target biomolecules. Adjust the sample conductivity close to that of the equilibration buffer; dialyze, desalt, or dilute high-salt samples.
Column Method
1. Column Packing and Equilibration: Thoroughly resuspend the Heparin Agarose 6FF, pack it into a suitable column, and equilibrate with at least 5 column volumes of equilibration/wash buffer.
2. Sample Loading: Load slowly using a pump, sample loop, or gravity flow and collect the flow-through. Do not exceed the binding capacity of the medium.
3. Washing: Wash with 5-10 column volumes of equilibration/wash buffer until the OD280 or UV absorbance reaches a stable baseline.
4. Elution: Elute with the NaCl-containing elution buffer using step or linear-gradient elution. Begin with a low salt concentration and increase gradually to 1-2 M NaCl while collecting fractions.
5. Regeneration: Remove residual bound material with a high-salt buffer, then restore the initial conditions using ddH2O and equilibration buffer. If compatible with the medium, mild alkaline cleaning may be used when required.
6. Storage: Equilibrate with storage buffer and store at 2-8°C. Do not freeze or dry.
Centrifugation Method
1. Resin Pretreatment: Transfer an appropriate amount of resin suspension into a centrifuge tube, centrifuge and discard the storage solution, and wash with 5 resin volumes of equilibration/wash buffer 2-3 times.
2. Binding: Add the prepared sample and incubate on a rotator at 4°C for 1-2 h or overnight.
3. Washing: Centrifuge, discard the supernatant, and wash with 5 resin volumes of equilibration/wash buffer 3-5 times.
4. Elution: Add 3-5 resin volumes of elution buffer, incubate at room temperature for 5-10 min, centrifuge, and collect the supernatant. Repeat 2-3 times if necessary.
Storage
4°C, 2 years
Do not dry or freeze
Attention
1. Ensure the resin is thoroughly resuspended before use.
2. It is recommended to replace the storage buffer every 2-3 months to prevent microbial contamination due to ethanol evaporation.
3. This product is for R&D use only, not for drug, household, or other uses.
4. For your safety and health, please wear a lab coat and disposable gloves to operate.
Components
| Components | HY-K0253-1 mL | HY-K0253-5 mL | HY-K0253-25 mL | HY-K0253-100 mL |
|---|---|---|---|---|
| Heparin Agarose 6FF | 1 mL | 5 mL | 25 mL | 100 mL |
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