products | Affinity Chromatography |
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Affinity Chromatography
Affinity Chromatography Columns and Matrices
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Phenylboronate columns |
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Thio-DVS Agarose Columns |
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Protein A columns |
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Protein G columns |
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DVS Agarose matrix |
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Aminophenyl-ATP-Sepharose® |
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O-Phospho-L-Tyrosine-Sepharose® |
Features:
- stable ligand immobilization
- high coupling capacity
- high mechanical stability
- improved flow characteristics
- re-usable
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| 5 ml column (left) and 2.5 ml Phenylboronate column (right). |
Product Description:
MoBiTec offers a variety of excellent affinity matrices with very stable ligand immobilization. The matrices provide high coupling capacity, high mechanical stability and improved flow characteristics. The individual ligands are coupled in an optimal concentration to the matrix. All matrices are packed into our unique 1 ml, 2.5 ml, 5 ml or 10 ml MoBiTec columns. They can be regenerated easily and be re-used many times.
| Matrix | Volume size | Purification of | Binding Capacity/Ligand Activity |
| Phenylboronate (PheBo) | 2 ml and 5 ml | beta-lactamase | >3 mg beta-lactamase/ml matrix |
| Protein A | 1 ml and 5 ml | immunoglobulins, polyclonal and monoclonal IgG | human polyclonal IgG: 39 mg/ml mouse monoclonal IgG 1: 11 mg/ml |
| Protein G | 1 ml and 5 ml | immunoglobulins, polyclonal and monoclonal IgG | human polyclonal IgG: >20 mg/ml matrix |
| Thio-DVS Agarose | 2 ml | immunoglobulins (IgG, IgM, IgA) and proteins (thio adsorption) | 10-50 mg/ml for proteins; 20-30 mg/ml for immunoglobulins |
| DVS Agarose | see below | ||
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Phenylboronate columns
Phenylboronate is a very stable matrix with affinity to beta-lactamase, Igase and peroxidase. Columns with immobilized phenylboronate specifically bind beta-lactamase in loading buffer; the enzyme can be washed out off the columns by elution buffer. The columns are particularly useful for the isolation of fusion proteins containing beta-lactamase, such as produced e.g. with our PheBo Fusion Protein Cloning System.
Columns properties:
Molecules of 3 x 107 Da and larger are excluded from the sepharoseTM beads. The columns can be re-used at least 10 times and can be stored for at least 5 months. About 10 mmole phenylboronate are immobilized per ml sepharoseTM beads. Following the general protocol, 1 ml of phenylboronate sepharoseTM binds at least 3 mg beta-lactamase.
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| The 2.5 ml column with Thio-DVS-Agarose (left) and Phenylboronate (middle) matrices. |
Thio-DVS Agarose Columns
Thio-DVS Agarose is a matrix with a low molecular weight ligand of non-protein origin. The thiophilic ligand is immobilized on Divinyl-Sulfon (DVS) activated agarose. Most proteins will bind to the new Thio-DVS-Agarose at high concentrations of ammonium sulfate and will easily be eluted by buffer. Neither organic solvents nor detergents are necessary for most proteins. Regeneration is very simple. The major applications include the purification of human, mouse and rabbit immunoglobulins, monoclonal antibodies and proteins as well as the concentration of dilute protein solutions.
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Protein A columns
CR-columns with immobilized Protein A may be used for many applications including: purification of monoclonal antibodies, purification or removal of polyclonal IgG from serum, or separation of IgG subclasses using a gradual pH gradient.
They are ideally suited for the efficient, cost effective and rapid purification of antibodies with minimal pre-treatment. The capacity for murine IgG at physiological pH and salt concentration are greater than that for other Protein A adsorbents and, in addition, the columns have very good flow properties.
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Protein G columns
In many instances antibodies bind more strongly to Protein G than to Protein A. Immobilized Protein G can be used for cost-effective purification of monoclonal and polyclonal antibodies which may be difficult or impossible to purify on immobilized Protein A columns, e.g. rat IgG2b and human IgG3. Protein G columns consist of recombinant Protein G immobilized on controlled pore glass which is more rigid and durable than conventional polymeric matrices.
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DVS Agarose matrix
An activated DVS Agarose Matrix is available for the immobilization of your biomolecules of choice, such as e.g. proteins or carbohydrates. For your convenience, we provide empty 2.5 ml columns and Mobicols (1 ml) with the matrix.
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Aminophenyl-ATP-Sepharose®, C10-spacer or w/o spacer
γ-Aminophenyl-adenosine triphospate (AP-ATP) immobilized on agarose, suitable for purifying ATP-binding proteins.
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Degree of substitution: 20 µmole AP-ATP/ml gel (C10-spacer); 25 µmol AP-ATP/ml gel (no spacer) Also available as prepacked column (AC-101CJB, AC-102CJB). |
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Selected references:
Haystead et al. (1993) Gamma-phosphate-linked ATP-Sepharose for the affinity purification of protein-kinases – rapid purification to homogeneity of sceletal-muscle mitogen-activated protein-kinase. Eur. J. Biochem.214:459.
Drewes et al. (1995) Microtubule-associated Protein/Microtubule Affinityregulating Kinase (p110 mark). J. Biol. Chem. 270: 7679.
The use of this reagent for the purification of protein kinases is subject to US patent No. 5.536.822. Such use is prohibited without a license from Serenex Inc. Information on obtaining a license is available at licensing@serenex.com.
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order information |
O-Phospho-L-Tyrosine-Sepharose®, C10-spacer or w/o spacer
O-Phospho-L-tyrosine immobilized on agarose, suitable for purification of proteins containing an SH2-domain.
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Degree of substitution: 25 µmole O-Phospho-L-tyrosine/ml gel Spacer: C10/none Storage buffer: 20% Also available as prepacked column (AC-103CJB, AC-104CJB). |
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Selected references:
Fry et al. (1992) Purification and characterization of a phosphatidyl 3- kinase complex from bovine brain by using phosphopeptide affinity columns. Biochem. J. 288:383.
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order information |
For a detailed data sheet as a pdf-file, please click on the product No.
| ORDER INFORMATION;SHIPPING & STORAGE: | ||
| order# | description | amount |
| PB0702 | 2 ml Phenylboronate column | 1 |
| PB0705 | 5 ml Phenylboronate column | 1 |
| PA0201 | 1 ml Protein A column | 1 |
| PA0205 | 1 ml Protein A column | 5 |
| PA0301 | 5 ml Protein A column | 1 |
| PS0501 | 1 ml Guard A column | 1 |
| PS0505 | 1 ml Guard A column | 5 |
| PS0601 | 5 ml Guard A column | 1 |
| PG0401 | 1 ml Protein G column | 1 |
| PG0402 | 1 ml Protein G column | 5 |
| PG0405 | 5 ml Protein G column | 1 |
| KEM940 | 2 ml Thio-DVS Agarose columns | 5 |
| KEM270 | 10 ml DVS Agarose matrix + empty columns (5 x 2.5 ml columns + 5 Mobicols) |
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| KEM275 | 50 ml DVS Agarose matrix + empty columns (10 x 2.5 ml columns + 10 Mobicols) |
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| AC-101SJB* | Aminophenyl-ATP Sepharose®, C10-spacer | 1 ml |
| AC-101LJB* | Aminophenyl-ATP Sepharose®, C10-spacer | 5 ml |
| AC-101CJB* | Aminophenyl-ATP Sepharose®, C10-spacer, 1 ml column | 1 |
| AC-102SJB* | Aminophenyl-ATP Sepharose®, no spacer | 1 ml |
| AC-102LJB* | Aminophenyl-ATP Sepharose®, no spacer | 5 ml |
| AC-102CJB* | Aminophenyl-ATP Sepharose®, no spacer, 1 ml column | 1 |
| AC-103SJB* | O-P-L-Tyrosine-Sepharose®, C10-spacer | 1 ml |
| AC-103LJB* | O-P-L-Tyrosine-Sepharose®, C10-spacer | 5 ml |
| AC-103CJB* | O-P-L-Tyrosine-Sepharose®, C10-spacer, 1 ml column | 1 |
| AC104SJB* | O-P-L-Tyrosine-Sepharose®, no -spacer | 1 ml |
| AC-104LJB* | O-P-L-Tyrosine-Sepharose®, no -spacer | 5 ml |
| AC-104CJB* | O-P-L-Tyrosine-Sepharose®, no -spacer, 1 ml column | 1 |
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shipped at RT; store at 4°C shipped on blue ice, store at 4°C |
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See also chapter 8 of our online catalog.
Download Handbooks:DVS Agarose, Thio-DVS Agarose & Protein G
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