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Columns for Bioseparation
| Selecting Columns for the Octave System
Some factors to consider when selecting columns for use with the Octave System... |
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The Semba Octave™ Chromatography System has a non-metallic flow path making it suitable for use with biological samples including:
- crude cell lysates
- tissue culture fluids
- serum samples
In all cases, as with any high performance chromatography system, samples should be free of particulate matter. Filtration of samples through a 0.45 micron membrane is usually sufficient.
The Semba Octave™ Chromatography System is an "open platform" permitting the use of a wide range of chromatography columns and solid phases. The system accommodates a variety of column sizes. In general, columns up to 2 cm x 10 cm packed with any polymeric resin (e.g. cross-linked agarose, methacrylate, polyacrylamide) having a particle size greater than 30 microns in diameter are compatible with the Octave System.
An important consideration for the type of column to be used with the Semba Octave System is the expected back pressure that will be generated during operation. The total fluid back pressure limit for the instrument is 270 psi (18.6 bar), which is due to the pressure limit of the pneumatic valve system (300 psi). Note that back pressures are additive. During typical applications no more than 3 columns are in any one zone, so in general if the back pressure generated by a single column with a given buffer and/or solvent and flow rate is less than 90 psi (6.2 bar), it should be compatible with the system.
Histidine-tagged Protein Purification - Octave™
Ni-NTA SF Columns:
See Semba's line of prepacked high-performance column sets for continuous, high-throughput purification of his-tagged proteins...
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The Octave™ Ni-NTA SF Columns are packed with the Ni-NTA Superflow, the most-cited resin for the purification of histidine-tagged recombinant proteins from any expression system under native or denaturing conditions. Ni-NTA Superflow provides Ni-NTA (nickel-nitrilotriacetic acid), coupled to Superflow resin and combines superior mechanical stability and outstanding flow characteristics with the high binding capacity of up to 50 mg/ml. NTA, which has four chelation sites for nickel ions, binds nickel more tightly than metal-chelating purification systems that only have three sites available for interaction with metal ions. The extra chelation site prevents nickel-ion leaching and results in a greater binding capacity and higher purity protein preparations.
The robust Superflow matrix allows flow rates up to 10 ml/min (1-ml columns) and 40 ml/min (5-ml columns), speeding the purification procedure and increasing throughput. The Octave Ni-NTA SF columns are compatible with a wide range of buffer components, including strong denaturants, detergents, and reducing agents. The columns are quickly and easily connected to the Octave Chromatography System through the 10-32 F to M6 F adapters (available separately).
Octave™ Ni-NTA SF Column Specs:
Binding capacity:
Up to 50 mg/ml (up to 2500 nmol @ ~20 kDa)
Support:
Superflow
Bead structure:
Highly cross-linked, 6% agarose
Bead size:
60–160 µm
Column dimensions, 1 ml:
6.7 mm I.D. x 28 mm L
Column dimensions, 5 ml:
14.7 mm I.D. x 29.8 mm L
Column housing material
Polypropylene
Cat. No.
Product Name
Price
C1001S
Octave™ Ni-NTA SF Column, 1 ml,
Set of 8*
$238.00
C1002S
Octave™ Ni-NTA SF Column, 5 ml,
Set of 8*
$790.00
A1015S
Adapter, 10-32 F to M6 F, Set of 8
$175.00
* Manufactured by Qiagen, Inc.
Product Literature
Technical Bulletin
MSDS
Certificate of Analysis
| Histidine-tagged Protein Purification - Octave™
Ni-NTA SF Columns: See Semba's line of prepacked high-performance column sets for continuous, high-throughput purification of his-tagged proteins... |
[−] |
The Octave™ Ni-NTA SF Columns are packed with the Ni-NTA Superflow, the most-cited resin for the purification of histidine-tagged recombinant proteins from any expression system under native or denaturing conditions. Ni-NTA Superflow provides Ni-NTA (nickel-nitrilotriacetic acid), coupled to Superflow resin and combines superior mechanical stability and outstanding flow characteristics with the high binding capacity of up to 50 mg/ml. NTA, which has four chelation sites for nickel ions, binds nickel more tightly than metal-chelating purification systems that only have three sites available for interaction with metal ions. The extra chelation site prevents nickel-ion leaching and results in a greater binding capacity and higher purity protein preparations.
The robust Superflow matrix allows flow rates up to 10 ml/min (1-ml columns) and 40 ml/min (5-ml columns), speeding the purification procedure and increasing throughput. The Octave Ni-NTA SF columns are compatible with a wide range of buffer components, including strong denaturants, detergents, and reducing agents. The columns are quickly and easily connected to the Octave Chromatography System through the 10-32 F to M6 F adapters (available separately).
| Octave™ Ni-NTA SF Column Specs: | |||
Binding capacity: |
Up to 50 mg/ml (up to 2500 nmol @ ~20 kDa) | ||
Support: |
Superflow | ||
Bead structure: |
Highly cross-linked, 6% agarose | ||
Bead size: |
60–160 µm | ||
Column dimensions, 1 ml: |
6.7 mm I.D. x 28 mm L | ||
Column dimensions, 5 ml: |
14.7 mm I.D. x 29.8 mm L | ||
| Column housing material | Polypropylene | ||
Cat. No. |
Product Name | Price | |||
C1001S |
Octave™ Ni-NTA SF Column, 1 ml, Set of 8* |
$238.00 | |
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C1002S |
Octave™ Ni-NTA SF Column, 5 ml, Set of 8* |
$790.00 | |
||
A1015S |
Adapter, 10-32 F to M6 F, Set of 8 | $175.00 | |
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| * Manufactured by Qiagen, Inc. | |||||
| Product Literature | |||||
| Technical Bulletin | |||||
| MSDS | |||||
| Certificate of Analysis | |||||
Imidazole, SembaChrom™ Grade, 2.5 M Solution:
The convenience and assurance of pure, low UV-absorbing, qualified & pH adjusted reagent for IMAC protein purification..
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Imidazole, SembaChrom™ Grade, is a special preparation of imidazole having low UV absorbance. Solutions containing imidazole are commonly used for the purification of histidine-tagged proteins by immobilized metal affinity chromatography (IMAC). Many commercial imidazole preparations contain UV-absorbing contaminants that interfere with spectrophotometric readings of protein solutions at 280 nm. SembaChrom Grade Imidazole has been processed to remove these contaminants, resulting in negligible background levels. The ready-to-use 2.5 M solution has been adjusted to pH 8.0 and filtered for convenient preparation of IMAC buffers.
Cat. No.
Product Name
Size
Price
R1002E
Imidazole, SembaChrom Grade,
2.5 M Solution
100 ml
$65.00
Product Literature
MSDS
Certificate of Analysis
| Imidazole, SembaChrom™ Grade, 2.5 M Solution: The convenience and assurance of pure, low UV-absorbing, qualified & pH adjusted reagent for IMAC protein purification.. |
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Imidazole, SembaChrom™ Grade, is a special preparation of imidazole having low UV absorbance. Solutions containing imidazole are commonly used for the purification of histidine-tagged proteins by immobilized metal affinity chromatography (IMAC). Many commercial imidazole preparations contain UV-absorbing contaminants that interfere with spectrophotometric readings of protein solutions at 280 nm. SembaChrom Grade Imidazole has been processed to remove these contaminants, resulting in negligible background levels. The ready-to-use 2.5 M solution has been adjusted to pH 8.0 and filtered for convenient preparation of IMAC buffers.
Cat. No. |
Product Name | Size | Price | ||
| R1002E | Imidazole, SembaChrom Grade, 2.5 M Solution |
100 ml | $65.00 | |
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| Product Literature | |||||
| MSDS | |||||
| Certificate of Analysis | |||||
| Antibody Purification: Octave™ MabCapture® A Columns See Semba's line of prepacked high-performance columns for rapid, high-throughput, antibody purification... |
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The Octave MabCapture A Columns are designed for rapid Protein A affinity purification of antibodies using Perfusion Chromatography® technology. The columns are available in 1-ml and 5-ml sizes pre-packed with POROS® MabCapture A media. The patented POROS polymer support enables use of much higher flow rates than conventional media without loss of binding capacity or resolution, resulting in faster separations. The rigid POROS particles are also less susceptible to compression and backpressure limitations even when operating at flow rates up to 4500 cm/h, permitting efficient operation over a broad range of conditions. The unique pore distribution and proprietary surface chemistry facilitate efficient coupling of Protein A ligand and unrivaled antibody binding capacity, with low leaching of Protein A through many cycles of binding and elution. The robust POROS matrix allows flow rates up to 20 ml/min (1-ml columns) and 80 ml/min (5-ml columns), speeding the purification procedure and increasing throughput. The columns are quickly and easily connected to the Octave Chromatography System through 10-32 F to M6 F adapters (available separately). The columns can also be used for automated purification on single-column liquid chromatography systems and for manual purification using a syringe.
The advantages of POROS MabCapture A include:
- The highest dynamic binding capacity for monoclonal antibodies vs. conventional agarose or silica based Protein A media
- Linear flow rates from 1000 cm/h to 4500 cm/h without loss of capacity or performance
- Compatibility with standard sodium hydroxide sanitation agents utilized for other chromatography products
- Perfect match with the Octave™ Chromatography System for ultimate efficiency and performance in preparative monoclonal antibody capture and purification
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| Octave MabCapture A Column Specs: | |||
Binding capacity: |
>45 mg/ml | ||
Support: |
Cross-linked Poly (styrene-divinylbenzene) | ||
Ligand: |
Recombinant Protein A | ||
Bead size: |
45 micron | ||
Column dimensions, 1 ml: |
6.7 mm I.D. x 28 mm L | ||
Column dimensions, 5 ml: |
14.7 mm I.D. x 29.8 mm L | ||
| Column housing material: | Polypropylene | ||
Cat. No. |
Product Name | Size | Price | ||
| C1007S | Octave™ MabCapture® A Columns | 8 x 1 ml | $695.00 | |
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| C1008S | Octave™ MabCapture® A Columns | 8 x 5 ml | $2350.00 | |
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| A1015S | Adapter, 10-32 F to M6 F, Set of 8 | set of 8 | $175.00 | |
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| C1009S | Octave™ MabCapture® A Columns | 2 x 1 ml | $190.00 | |
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| C1008E | Octave™ MabCapture® A Columns | 1 x 5 ml | $345.00 | |
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| A1018S | Adapter, 10-32 F to M6 F, Set of 2 | set of 2 | $44 | |
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| Made with POROS® MabCapture® A media from Life Technologies Corporation. POROS® and MabCapture® are Registered Trademarks of Life Technologies Corporation | |||||
| Product Literature | |||||
| Technical Bulletin | |||||
| MSDS | |||||
| Certificate of Analysis | |||||