GST-Sepharose 4B Gravity Prepacked Column
| 【Numbering】BK-NRPB56S | 【CAS】CAS | ||
| 【Item No.】BK-NRPB56S-Z25 | 【specification】 | ||
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Product name: GST-Sepharose 4B
English name: Glutathione NUPharose 4B
Specifications: 20 ml, 100 ml, 1L, 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 4B is a bio-affinity chromatography separation medium formed by bonding glutathione to agarose gel microspheres. It is mainly used for glutathione transferase-labeled protein (GST fusion protein). ), glutathione transferase and glutathione-dependent protein separation and purification, through one-step affinity chromatography, high-purity protein can be obtained. Compared with GST-Sepharose FF, GST-Sepharose 4B can obtain a higher load.
Technical index:
Matrix
4% highly cross-linked agarose gel
Ligand
Glutathione
Ligand Density
≥20μmol/ml
Filler particle size
60~180 μm
Protein load
About 25mg glutathione-S-transferase (28kDa)/ml
Maximum flow rate
100 cm/h
Recommended flow rate
10~50 cm/h
Back pressure resistant
0.1 MPa
pH stability
3-12 (long time); 3-13 (short time)
Chemical stability
Stable in the following solutions: commonly used aqueous buffer; 0.1mol/L NaOH, 6mol/L guanidine hydrochloride; 70% ethanol.
Instructions:
1. Pack the column
1.1 Prepare the initial buffer (equilibration 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, make a homogenate with distilled water and degas.
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 to pour into the column along the inner wall of the column at one time, so that the gel can 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 2-3 times the column volume of buffer.
Note: (1) Air bubbles cannot be introduced during all operations to ensure the uniformity of the glue filling. (3) 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. (4) 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: 10mM PBS (140mM NaCl, 2.7mM KCl, 10mM Na2HPO4, 1.8mM KH2PO4, pH 7.4).
3. 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
Rinse the loaded chromatography column with 2-3 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 10mM reduced glutathione solution (may contain buffer components) for elution. The recommended elution buffer is: 50mM Tris-HCl, 10 mM reduced glutathione (GSH), pH 8.0.
6. Regeneration
Flush 3-5 column volumes with distilled water at the operating flow rate, and then wash with balance solution to balance 3-5 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)
Remove precipitates and denatured proteins: wash with 2CV 6M guanidine hydrochloride, and immediately wash 5CV with 10mM PBS (pH7.4).
Remove strong hydrophobin: Wash with 3-4CV 70% ethanol, and immediately wash 5CV with 10mM PBS (pH7.4).
Message
English name: Glutathione NUPharose 4B
Specifications: 20 ml, 100 ml, 1L, 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 4B is a bio-affinity chromatography separation medium formed by bonding glutathione to agarose gel microspheres. It is mainly used for glutathione transferase-labeled protein (GST fusion protein). ), glutathione transferase and glutathione-dependent protein separation and purification, through one-step affinity chromatography, high-purity protein can be obtained. Compared with GST-Sepharose FF, GST-Sepharose 4B can obtain a higher load.
Technical index:
Matrix
4% highly cross-linked agarose gel
Ligand
Glutathione
Ligand Density
≥20μmol/ml
Filler particle size
60~180 μm
Protein load
About 25mg glutathione-S-transferase (28kDa)/ml
Maximum flow rate
100 cm/h
Recommended flow rate
10~50 cm/h
Back pressure resistant
0.1 MPa
pH stability
3-12 (long time); 3-13 (short time)
Chemical stability
Stable in the following solutions: commonly used aqueous buffer; 0.1mol/L NaOH, 6mol/L guanidine hydrochloride; 70% ethanol.
Instructions:
1. Pack the column
1.1 Prepare the initial buffer (equilibration 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, make a homogenate with distilled water and degas.
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 to pour into the column along the inner wall of the column at one time, so that the gel can 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 2-3 times the column volume of buffer.
Note: (1) Air bubbles cannot be introduced during all operations to ensure the uniformity of the glue filling. (3) 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. (4) 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: 10mM PBS (140mM NaCl, 2.7mM KCl, 10mM Na2HPO4, 1.8mM KH2PO4, pH 7.4).
3. 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
Rinse the loaded chromatography column with 2-3 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 10mM reduced glutathione solution (may contain buffer components) for elution. The recommended elution buffer is: 50mM Tris-HCl, 10 mM reduced glutathione (GSH), pH 8.0.
6. Regeneration
Flush 3-5 column volumes with distilled water at the operating flow rate, and then wash with balance solution to balance 3-5 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)
Remove precipitates and denatured proteins: wash with 2CV 6M guanidine hydrochloride, and immediately wash 5CV with 10mM PBS (pH7.4).
Remove strong hydrophobin: Wash with 3-4CV 70% ethanol, and immediately wash 5CV with 10mM PBS (pH7.4).
| 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 |
Message
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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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