GST-Sepharose FF medium pressure prepacked column
| 【Numbering】BK-NRPB08S | 【CAS】CAS | ||
| 【Item No.】BK-NRPB08S-Y25 | 【specification】 | ||
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Product name: GST-Sepharose FF
English name: Glutathione NUPharose Fast Flow, GST NUPharose FF
Specifications: 20 ml, 100 ml, 1 L, 1 ml pre-packed column, 5 ml pre-packed column, special specifications customized
Transportation: 2 ~ 30℃, normal pressure, avoid light
Storage: 20% ethanol, 2 ~ 8℃
Shelf life: 3 years
Related introduction:
GST-Sepharose FF is a bio-affinity chromatography separation medium formed by bonding glutathione to agarose gel microspheres. The product retains the excellent hydrophilicity of agarose and the large grid structure, and has good compatibility with biologically active macromolecules. It has the characteristics of high loading capacity, low non-specific adsorption, and fast flow rate. It is mainly used for grains. Separation and purification of glutathione transferase labeled protein (GST fusion protein), glutathione transferase and glutathione-dependent proteins.
Technical index:
Matrix
4% highly cross-linked agarose gel
Ligand
Glutathione
Ligand Density
≥10 μmol/ml
Filler particle size
60 ~ 180 μm
Protein load
About 10mg glutathione-S-transferase/mL
Maximum flow rate
300 cm/h
Recommended flow rate
20 ~ 100 cm/h
Back pressure resistant
0.3 MPa
pH stability
3 ~ 12 (long time); 3 ~ 13 (short time)
Chemical stability
Stable in the following solutions: commonly used aqueous buffers; 1 M acetic acid (pH 4.0); 6 M guanidine hydrochloride; 70% ethanol.
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 conductivity, pH and other parameters remain unchanged. The recommended binding equilibration buffer is: 10 mM PBS (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, pH 7.4).
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 chromatographic medium. Linear experiments 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 chromatography column with 2 to 3 column volumes of equilibration buffer and observe the changes of the detector until the conductivity, pH and other parameters remain unchanged, at which time the unexchanged components are washed out.
5. Elution
It is generally recommended to use 10 mM reduced glutathione solution (may contain buffer components) for elution. The recommended elution buffer is: 50 mM Tris-HCl, 10 mM reduced glutathione (GSH), 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)
To remove precipitates and denatured proteins: wash with 2 CV 6 M guanidine hydrochloride, and immediately wash with 10 mM PBS (pH 7.4) for 5 CV.
Removal of strong hydrophobin: Wash with 3 ~ 4 CV 70% ethanol, and immediately wash with 10 mM PBS (pH 7.4) for 5 CV.
Precautions:
Recommended flow rate
Pre-packed column volume (ml)
1 ml
5 ml
10 ml
Flow rate (ml /min)
0.2
1
2
Message
English name: Glutathione NUPharose Fast Flow, GST NUPharose FF
Specifications: 20 ml, 100 ml, 1 L, 1 ml pre-packed column, 5 ml pre-packed column, special specifications customized
Transportation: 2 ~ 30℃, normal pressure, avoid light
Storage: 20% ethanol, 2 ~ 8℃
Shelf life: 3 years
Related introduction:
GST-Sepharose FF is a bio-affinity chromatography separation medium formed by bonding glutathione to agarose gel microspheres. The product retains the excellent hydrophilicity of agarose and the large grid structure, and has good compatibility with biologically active macromolecules. It has the characteristics of high loading capacity, low non-specific adsorption, and fast flow rate. It is mainly used for grains. Separation and purification of glutathione transferase labeled protein (GST fusion protein), glutathione transferase and glutathione-dependent proteins.
Technical index:
Matrix
4% highly cross-linked agarose gel
Ligand
Glutathione
Ligand Density
≥10 μmol/ml
Filler particle size
60 ~ 180 μm
Protein load
About 10mg glutathione-S-transferase/mL
Maximum flow rate
300 cm/h
Recommended flow rate
20 ~ 100 cm/h
Back pressure resistant
0.3 MPa
pH stability
3 ~ 12 (long time); 3 ~ 13 (short time)
Chemical stability
Stable in the following solutions: commonly used aqueous buffers; 1 M acetic acid (pH 4.0); 6 M guanidine hydrochloride; 70% ethanol.
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 conductivity, pH and other parameters remain unchanged. The recommended binding equilibration buffer is: 10 mM PBS (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, pH 7.4).
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 chromatographic medium. Linear experiments 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 chromatography column with 2 to 3 column volumes of equilibration buffer and observe the changes of the detector until the conductivity, pH and other parameters remain unchanged, at which time the unexchanged components are washed out.
5. Elution
It is generally recommended to use 10 mM reduced glutathione solution (may contain buffer components) for elution. The recommended elution buffer is: 50 mM Tris-HCl, 10 mM reduced glutathione (GSH), 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)
To remove precipitates and denatured proteins: wash with 2 CV 6 M guanidine hydrochloride, and immediately wash with 10 mM PBS (pH 7.4) for 5 CV.
Removal of strong hydrophobin: Wash with 3 ~ 4 CV 70% ethanol, and immediately wash with 10 mM PBS (pH 7.4) for 5 CV.
Precautions:
Recommended flow rate
Pre-packed column volume (ml)
1 ml
5 ml
10 ml
Flow rate (ml /min)
0.2
1
2
| 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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