Heparin-Sepharose FF Gravity Prepacked Column
| 【Numbering】BK-NRPB09S | 【CAS】CAS | ||
| 【Item No.】BK-NRPB09S-Z25 | 【specification】 | ||
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Product name: Heparin-Sepharose FF
English name: Heparin NUPharose Fast Flow, Heparin NUPharose FF
Specifications: 20 ml, 100 ml, 1 L, 1 ml pre-packed column, 5 ml pre-packed column, special specifications customized
Transportation: 2 ~ 25℃, normal pressure, avoid light
Storage: 0.05M NaAc in 20% ethanol, 2 ~ 8℃
Shelf life: 3 years
Related introduction:
Heparin-Sepharose FF is a bioaffinity chromatography separation medium formed by bonding heparin to agarose gel microspheres. The product retains the excellent hydrophilicity of agarose and the large network structure, and has good compatibility with biologically active macromolecules. It has the characteristics of high load, low non-specific adsorption and fast flow rate. It is mainly used for blood coagulation. Separation and purification of enzyme III, coagulation factors, lipoproteins, esterases, hormones, interferons, etc.
Technical index:
Matrix
6% cross-linked agarose
Ligand
heparin
Ligand Density
5 mg/mL
Protein load
About 2 mg antithrombin Ⅲ /mL
Maximum flow rate
800 cm/h
Recommended flow rate
20 ~ 100 cm/h
Back pressure resistant
0.3 MPa
Particle size range
60 ~ 180 μm
Operating temperature
4 ~ 40℃
pH stability
4 ~ 12 (long time); 3 ~ 14 (short time)
Heat resistant
Sterilize in pH 7 water at 120°C for 30 min
Instructions:
1. Column loading
1.1 Prepare initial buffer (equilibrium solution) and elution buffer according to the nature of the separation target.
1.2 Drain the gel and wash it twice with distilled water to remove the stored ethanol, mix it with distilled water and degas it.
1.3 Fix the column vertically, moisten the bottom end with water or buffer and keep the liquid level for a period of time.
1.4 Use a glass rod to guide the homogenate and pour it into the column along the inner wall of the column at one time to allow the gel to settle freely in the column.
1.5 Connect the movable column head at the top of the column, turn on the peristaltic pump, let the buffer flow through 5 column volumes at the operating flow rate during use, and then use 1.5 times the operating flow rate to flow through 5 column volumes, adjust the adapter column head to make it as close as possible to the glue surface , And finally equilibrate the column with 2 to 3 column volumes of buffer.
Notice:
1) No air bubbles can be introduced during all operations to ensure the uniformity of the glue filling.
2) If 1.2 is done unconditionally, there are bubbles in the packing layer, and the column can be loaded twice to remove the stored ethanol and bubbles.
3) The solution prepared by reagents such as ethanol needs to be degassed.
2. Balance
Equilibrate the column with the equilibration buffer at the operating flow rate, and observe the changes in the detector until the parameters such as UV, pH, and conductivity remain unchanged. The recommended equilibration buffer is: 50 mM Tris-HCl, pH 8.0.
3. Sample loading
Switch the switching valve to load the sample. The sample amount is selected according to the nature of the sample and the amount of the chromatography medium. The linear experiment can also be performed to find the best sample amount; sample pretreatment: replacement of buffer, clarification and filtration (0.45, 0.22) μm) and so on.
4. Rinse
Wash the loaded chromatographic column with 2 to 3 column volumes of equilibration buffer and observe the changes of the detector until the parameters such as UV, pH, and conductivity remain unchanged.
5. Elution
It is generally recommended to use a high-salt solution (which may contain buffer components) for elution. In order to obtain better purity, it can be eluted in a gradient. The recommended elution buffer is: 50 mM Tris-HCl, 2 M NaCl, pH 8.0.
6. Regeneration
Wash 3 to 5 column volumes with distilled water at the operating flow rate, and then wash with a balance solution to balance 3 to 5 column volumes.
If there are inactivated proteins or lipids that cannot be washed off during regeneration, they can be removed by cleaning-in-place (CIP).
7. Cleaning-in-place (CIP)
For strong binding proteins or lipids, the following process can be used for in-place cleaning: 0.1 mol/L NaOH washes 2 column volumes, 8 mol/L urea washes 2 column volumes, and equilibration buffer washes 2 column volumes.
Message
English name: Heparin NUPharose Fast Flow, Heparin NUPharose FF
Specifications: 20 ml, 100 ml, 1 L, 1 ml pre-packed column, 5 ml pre-packed column, special specifications customized
Transportation: 2 ~ 25℃, normal pressure, avoid light
Storage: 0.05M NaAc in 20% ethanol, 2 ~ 8℃
Shelf life: 3 years
Related introduction:
Heparin-Sepharose FF is a bioaffinity chromatography separation medium formed by bonding heparin to agarose gel microspheres. The product retains the excellent hydrophilicity of agarose and the large network structure, and has good compatibility with biologically active macromolecules. It has the characteristics of high load, low non-specific adsorption and fast flow rate. It is mainly used for blood coagulation. Separation and purification of enzyme III, coagulation factors, lipoproteins, esterases, hormones, interferons, etc.
Technical index:
Matrix
6% cross-linked agarose
Ligand
heparin
Ligand Density
5 mg/mL
Protein load
About 2 mg antithrombin Ⅲ /mL
Maximum flow rate
800 cm/h
Recommended flow rate
20 ~ 100 cm/h
Back pressure resistant
0.3 MPa
Particle size range
60 ~ 180 μm
Operating temperature
4 ~ 40℃
pH stability
4 ~ 12 (long time); 3 ~ 14 (short time)
Heat resistant
Sterilize in pH 7 water at 120°C for 30 min
Instructions:
1. Column loading
1.1 Prepare initial buffer (equilibrium solution) and elution buffer according to the nature of the separation target.
1.2 Drain the gel and wash it twice with distilled water to remove the stored ethanol, mix it with distilled water and degas it.
1.3 Fix the column vertically, moisten the bottom end with water or buffer and keep the liquid level for a period of time.
1.4 Use a glass rod to guide the homogenate and pour it into the column along the inner wall of the column at one time to allow the gel to settle freely in the column.
1.5 Connect the movable column head at the top of the column, turn on the peristaltic pump, let the buffer flow through 5 column volumes at the operating flow rate during use, and then use 1.5 times the operating flow rate to flow through 5 column volumes, adjust the adapter column head to make it as close as possible to the glue surface , And finally equilibrate the column with 2 to 3 column volumes of buffer.
Notice:
1) No air bubbles can be introduced during all operations to ensure the uniformity of the glue filling.
2) If 1.2 is done unconditionally, there are bubbles in the packing layer, and the column can be loaded twice to remove the stored ethanol and bubbles.
3) The solution prepared by reagents such as ethanol needs to be degassed.
2. Balance
Equilibrate the column with the equilibration buffer at the operating flow rate, and observe the changes in the detector until the parameters such as UV, pH, and conductivity remain unchanged. The recommended equilibration buffer is: 50 mM Tris-HCl, pH 8.0.
3. Sample loading
Switch the switching valve to load the sample. The sample amount is selected according to the nature of the sample and the amount of the chromatography medium. The linear experiment can also be performed to find the best sample amount; sample pretreatment: replacement of buffer, clarification and filtration (0.45, 0.22) μm) and so on.
4. Rinse
Wash the loaded chromatographic column with 2 to 3 column volumes of equilibration buffer and observe the changes of the detector until the parameters such as UV, pH, and conductivity remain unchanged.
5. Elution
It is generally recommended to use a high-salt solution (which may contain buffer components) for elution. In order to obtain better purity, it can be eluted in a gradient. The recommended elution buffer is: 50 mM Tris-HCl, 2 M NaCl, pH 8.0.
6. Regeneration
Wash 3 to 5 column volumes with distilled water at the operating flow rate, and then wash with a balance solution to balance 3 to 5 column volumes.
If there are inactivated proteins or lipids that cannot be washed off during regeneration, they can be removed by cleaning-in-place (CIP).
7. Cleaning-in-place (CIP)
For strong binding proteins or lipids, the following process can be used for in-place cleaning: 0.1 mol/L NaOH washes 2 column volumes, 8 mol/L urea washes 2 column volumes, and equilibration buffer washes 2 column volumes.
| Warm tips: Suzhou Beike nano products are only used for scientific research, not for human body,different batches of products have different specifications and performance |
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| Warm tips: Suzhou Beike nano products are only used for scientific research, not for human body,different batches of products have different specifications and performance.The website pictures are from the Internet. The pictures are for reference only. Please take the real object as the standard. In case of infringement, please contact us to delete them immediately. |
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