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HisCap 6FF
Ni NTA Beads 6FF is based on a highly cross-linked 6% agarose with the same ligand as Ni NTA Beads (See Fig 1.), see table 1. In addition to being able to tolerate harsh reagent conditions (see Table 2), Ni NTA Beads 6FF is more stable due to its pressure-resistant matrix, which can withstand pressures up to 0.3 MPa, making the product is more suitable for industrial large-scale protein purification, which can be achieved at relatively high flow rates for the purification of target proteins. HisCap 6FF is one of a range of prepacked, ready-to-use columns for affinity chromatography. It is packed with 1ml and 5ml of Ni NTA Beads 6FF. Five different packing sizes are available. HisCap 6FF has the standard interface and can be adapted to all kinds of chromatography system, such as ÄKTA. It is fast, simple and easy operation.
· Product Description
Table 1. Characteristics of Ni NTA Beads 6FF
| Item | Description |
|---|---|
| Matrix Spherical | Highly cross-linked 6% agarose |
| Static Binding Capacity | >40 mg 6×His-tagged protein/ml medium |
| Particle size | 45-165 μm |
| Maximum Pressure | 0.3 MPa, 3 bar |
| Storage Solution | 1×PBS containing 20% ethanol |
| Storage Temperature | 4-30°C |
Table 2. Chemical compatibilities for Ni NTA Beads 6FF
| Reagent | Stability |
|---|---|
| Reductants | 5 mM DTE |
| 1mM DTT | |
| 20 mM β-mercaptoethanol | |
| 5 mM TCEP | |
| 10 mM reduced glutathione | |
| Denaturants | 8 M urea |
| 6 M Gua-HCl | |
| Detergent | 2% Triton™ X-100 (nonionic) |
| 2% Tween™ 20 (nonionic) | |
| 2% NP-40 (nonionic) | |
| 2% cholate (anionic) | |
| 1% CHAPS (zwitterionic) | |
| Other additives | 500 mM imidazole |
| 20% ethanol | |
| 50% glycerol | |
| 100 mM Na2SO4 | |
| 1.5 M NaCl | |
| 1 mM EDTA | |
| 60 mM citrate |
Table 3. Recommended buffer for his-tagged protein purification under native conditions
| Name | Volume | Ingredient |
|---|---|---|
| Lysis Buffer | 1 L | 50 mM NaH2PO4 (7.80 g NaH2PO4·2H2O) 300 mM NaCl (17.54 g NaCl) 10 mM imidazole (0.68 g imidazole) Adjust the buffer pH to 8.0 with NaOH solution. |
| Wash Buffer | 1 L | 50 mM NaH2PO4 (7.80 g NaH2PO4·2H2O) 300 mM NaCl (17.54 g NaCl) 20 mM imidazole (1.36 g imidazole) Adjust the buffer pH to 8.0 with NaOH solution. |
| Elution Buffer | 1 L | 50 mM NaH2PO4 (7.80 g NaH2PO4·2H2O) 300 mM NaCl (17.54 g NaCl) 250 mM imidazole (17.0 g imidazole) Adjust the buffer pH to 8.0 with NaOH solution. |
Table 4. Recommended buffer for his-tagged protein purification under denaturing conditions, pH elution
| Name | Volume | Ingredient |
|---|---|---|
| Lysis Buffer | 1 L | 8 M Urea (480.50 g Urea) 100 mM NaH2PO4 (15.60 g NaH2PO4·2H2O) 100 mM Tris·HCl (15.76 g Tris·HCl) Adjust the buffer pH to 8.0 with HCl solution |
| Wash Buffer | 1 L | 8 M Urea (480.50 g Urea) 100 mM NaH2PO4 (15.60 g NaH2PO4·2H2O) 100 mM Tris·HCl (15.76 g Tris·HCl) Adjust the buffer pH to 6.3 with HCl solution |
| Name | Volume | Ingredient |
| Elution Buffer | 1 L | 8 M Urea (480.50 g Urea) 100 mM NaH2PO4 (15.60 g NaH2PO4·2H2O) 100 mM Tris·HCl (15.76 g Tris·HCl) Adjust the buffer pH to 4.5 with HCl solution |
Table 5. Recommended buffer for his-tagged protein purification under denaturing conditions, imidazole elution
| Name | Volume | Ingredient |
|---|---|---|
| Lysis Buffer | 1 L | 8 M Urea (480.50 g Urea) 50 mM NaH2PO4 (7.80 g NaH2PO4·2H2O) 300 mM NaCl (17.54 g NaCl) 10 mM imidazole (0.68 g imidazole) Adjust the buffer pH to 8.0 with NaOH solution. |
| Wash Buffer | 1 L | 8 M Urea (480.50 g Urea) 50 mM NaH2PO4 (7.80 g NaH2PO4·2H2O) 300 mM NaCl (17.54 g NaCl) 20 mM imidazole (1.36 g imidazole) Adjust the buffer pH to 8.0 with NaOH solution. |
| Elution Buffer | 1 L | 8 M Urea (480.50 g Urea) 50 mM NaH2PO4 (7.80 g NaH2PO4·2H2O) 300 mM NaCl (17.54 g NaCl) 250 mM imidazole (17.0 g imidazole) Adjust the buffer pH to 8.0 with NaOH solution. |
Note: 8 M Urea could also be replaced by 6 M Guanidine-HCl. Table 4 or Table 5 select one of them as the solution for the inclusion body purification.
· Ordering Information
| Product | Cat. No. | Size |
|---|---|---|
| Ni NTA Beads | SA004005 | 5 ml |
| SA004025 | 25 ml | |
| SA004100 | 100 ml | |
| SA004500 | 500 ml | |
| SA00401L | 1 L | |
| SA00410L | 10 L | |
| HisPur Ni NTA Kit | SA004K03 | 3 times |
| Ni NTA Beads 6FF | SA005005 | 5 ml |
| SA005025 | 25 ml | |
| SA005100 | 100 ml | |
| SA005500 | 500 ml | |
| SA00501L | 1 L | |
| SA00510L | 10 L | |
| HisCap 6FF | SA005C11 | 1×1 ml |
| SA005C51 | 5×1 ml | |
| SA005C15 | 1×5 ml | |
| SA005C55 | 5×5 ml | |
| SA005CS | 3×1 ml+1×5 ml |
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