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 | 15 to 30°C |  | Ambient or blue ice | | Note: Store and Ship conditions may differ. | | See Key |
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| | Luer-lock Syringe Adaptor Set (for HP Cartridges) |
|  | Cat. No. 71769 |
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| Complete range of reagents and kits for extraction and purification | | | Affinity Purification
Affinity purification is based on the specific interaction of a target molecule with an immobilized ligand. Affinity technology can be used to isolate specific molecules from a mixture (e.g., His•Bind® purification), capture a desired molecule for interaction studies (e.g., immobilization on GST•Bind™ Resin), or remove a component from a reaction (e.g., protease removal with the Cleavage Capture Kits). For recombinant proteins, the addition of fusion tags using appropriate expression vectors enables affinity purification by a number of strategies. Our selection of immobilized metal affinity chromatography (IMAC) matrices for purification of His•Tag® fusion proteins is the widest available, as shown in the tables here and on the following two pages. Other agarose-based affinity resins allow rapid purification of GST•Tag™, S•Tag™, Strep•Tag® II, and T7•Tag® fusion proteins.
For the convenient, reproducible preparation of custom affinity supports using antibodies, peptides, proteins, or other user-defined ligands, Novagen offers an activated matrix ready for coupling. PreACT™ Agarose ALD is a preparation of aldehyde-activated cross-linked agarose that forms stable amide linkages with free amino groups of proteins or peptides in the presence of sodium cyanoborohydride.
Streptavidin Agarose and MagPrep® Streptavidin Beads provide a choice of formats for purification of biotin-containing molecules. Both supports are qualified for high biotin binding activity and low non-specific background, and are well-suited to a variety of applications that make use of the extraordinary affinity of biotin for streptavidin.
The MagPrep Anti-Ig Beads consist of a super-paramagnetic iron oxide core encapsulated within a polystyrene shell coated with antibodies against mouse, rat, or human IgG or IgM. These beads offer many advantages for highly selective capture and immobilization of antibodies and immune complexes. Protein A and Protein G agaroses are indispensible tools for affinity purification and immunoprecipitation of antibodies and their bound antigens. Two types of supports are available that contain highly purified Protein A and/or Protein G Plus, designed for purification or for immunoprecipitation, respectively.
Group separation and proteome complexity reduction kits
One of the major challenges in functional proteomics is the separation of complex protein mixtures to allow detection of low-abundance proteins and enrich for functional protein groups to facilitate downstream analysis. To help you meet this challenge, we offer the ProteoEnrich™ and ProteoExtract® product lines. The ProteoEnrich ATP-Binders kit allows group separation of protein kinases and other ATP-binding proteins, yielding partially purified cell extracts enriched in active protein kinases. Based on an affinity resin that contains immobilized ATP, the method is compatible with 2D electrophoresis, SDS-PAGE/tandem mass spectrometry, Western blot analysis, and activity assays.
The Novagen ProteoEnrich CAT-X and CAT-X SEC Kits provide highly specific, reproducible, and convenient methods to reduce proteome complexity, maximizing the likelihood to visualize low-abundance proteins. Through selective fractionation based on total protein charge or protein charge and molecular weight, the kits generate subproteome fractions compatible with downstream mass spectrometry, SELDI, or array-based analysis.
The ProteoExtract Phosphopeptide Capture Kit is a tool for phosphoproteome analysis that relies on the specific interaction of phosphate groups with immobilized zirconium ions on the surface of magnetic particles to capture highly purified phosphopeptides for identification by LC-MS or MALDI-MS. The ProteoExtract Phosphoproteome Profiler Kit provides a method to determine protein phosphorylation patterns resulting from external and internal signaling by selectively enriching the phosphorylated peptides through affinity purification on magnetic particles.
For additional information, see also our chapter on Protein Extraction, which highlights the ProteoExtract Extraction Kits, Abundant Protein Removal Kits, All-in-One Trypsin Digestion Kit, the Protein Precipitation Kit, and Detergent Set.
Separation Devices
For optimal performance and convenience, Novagen offers a range of devices for use with corresponding affinity resins. Three different Magnetight™ magnetic separation devices are available, designed for use with 96-well plates, multiple 1.5-ml centrifuge tubes, and 15- or 50-ml tubes. For devices using Luer Lock fittings, the Vacuum Manifold allows simultaneous vacuum processing of up to 12 cartridges and/or columns and is compatible with His•Bind Quick Cartridges and Columns.
Site-specific Proteases and Cleavage Capture Kits
Restriction grade site-specific proteases (Thrombin, biotinylated Thrombin, Factor Xa, HRV 3C Proease, Tag•off™ High Activity rEK, and recombinant Enterokinase) and Cleavage Capture Kits offer convenient removal of fusion tags. EKapture™ and Xarrest™ Agaroses are used for the quantitative removal of rEKs and Factor Xa, respectively, following cleavage of fusion proteins. Biotinylated thrombin is removed with Streptavidin Agarose and HRV 3C is removed with Ni-NTA agarose.
Protein Refolding
Overexpression of recombinant proteins in E.coli is an efficient method for producing large quantities of target proteins for many biological and new drug development applications. However, proteins often accumulate in the cells as inactive, misfolded, and insoluble aggregates (inclusion bodies).
Some purification strategies take advantage of the formation of insoluble inclusion bodies, which can sequester the target protein and protect it from degradation in the cell. To obtain the target protein in an active, soluble form, the inclusion bodies must be solubilized and the protein refolded. Unfortunately, defining conditions that promote refolding of the target protein into its native conformation is empirical and time consuming.
While the optimal conditions for solubilization and refolding largely depend on the individual protein, some general protocols and reagents have been successfully used for a number of different proteins. The Protein Refolding Kit provides the reagents to perform one general refolding protocol and can serve as a basis for developing an optimized refolding buffer, containing additional reagents such as non-detergent sulfobetaines (NDSBs, see page 310).
To determine optimal target protein refolding conditions, the iFOLD™ Protein Refolding System 1 includes all reagents needed to isolate and denature inclusion bodies, and a 96-well plate containing 92 unique refolding buffers. Optimal refolding conditions for target proteins are determined by systematically and simultaneously evaluating the 92 different refolding buffer combinations of pH, salt, cyclodextrin, redox agent, and refolding additives.
Dialysis Devices
D-Tube™ Dialyzers can be used for dialysis or electroelution of proteins or nucleic acids from polyacrylamide or agarose gels. The Electroelution Accessory Kit provides a compatible support tray and precipitation reagents.
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The Luer-lock Syringe Adaptors are designed for use with the Strep • Tactin ® Superflow™ HP Cartridges to provide compatibility with a Luer-lock syringe. The adaptor set contains 3 x female luer to 10-32 male adaptors and 3 x 1/16 inch barb to 10-32 male adaptors. |
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| Need additional information about this product? Email our Technical Service department at: novatech@novagen.com |
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Merck Chemicals USD list price is displayed (pricing with local distributors may vary). NOTE: In Stock status is based on item availability worldwide.
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