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| | | ProPac IMAC Columns |  | The ProPac IMAC-10 is a high-resolution HPLC column for the separation of proteins and peptides by immobilized metal affinity chromatography State-of-the-art technology for reusable columns with tailored specificity Resolve the target proteins using a single column in a single run Retention control by imidazole or pH gradient High purity separations of metal-binding proteins Wide range of metal-specific applications
IMAC is typically carried out using a capture/release method for protein and peptide enrichment. Often, the fractions collected contain impurities and further purification is required by reversed-phase HPLC, ion exchange or size exclusion. The ProPac IMAC-10 analytical column was developed as a high resolution HPLC column, to allow researchers to run linear gradient separations in the IMAC mode, resulting in highly efficient resolution of proteins and highly pure fractions collected. The recommended conditions for eluting bound protein from a Dionex ProPac IMAC-10 column is a linear imidazole gradient, because under these conditions it is possible to not simply elute the bound protein, but in some cases also to separate different bound fractions at different imidazole concentrations. The ProPac IMAC-10 column comes in a variety of dimensions and can be applied to a broad range of applications, including the separation of His-tagged protein aggregation variants, His-tagged protein purification, polishing samples for crystallization experiments, on-column refolding, phosphopeptide analysis, intact protein separations, separation of monoclonal antibodies and prion peptide analysis. The ProPac IMAC-10 phase consists of 10-µm, nonporous, polymeric beads which are coated with a hydrophilic layer to eliminate secondary hydrophobic interactions with the proteins. Poly(IDA) chains are then grafted onto the hydrophilic layer using state of the art technology. The poly(IDA) grafts are converted to metal containing nanoparticles when the column is charged with metal. It is these nanoparticles that act as the IMAC interaction sites for individual proteins and provide the ProPac IMAC-10 with it’s high resolving power. This design maximizes the number of sterically accessible ligands for binding to the proteins, thereby ensuring that the proteins remain tightly bound during the separation step. The ProPac IMAC-10 is shipped free from any metal and ready to be charged with the metal of your choice. It can be operated between pH 2-12, it is compatible with most reagents commonly used in protein purifications, such as denaturants, non-ionic detergents and reducing agents and it can be used under native or denaturing HPLC conditions.
| | ProPac IMAC Columns Specifications | | Maximum Operating Pressure | < 3,000 psi (XX MPa) | | Temperature Range | <60oC Recommended Temperature: Controlled ambient | | Typical Operating Conditions | <600 psi at 1.0 mL/min (4 x 250mm) Recommended flow rate: 1.0 mL/min | | Mobile Phase Compatibility | pH 2-12, Compatible with most reagents used in IMAC protein purifications, including EtOH, Urea, NaCl, non-ionic detergents, glycerol, acetic acid, guanidine, HCl | | Resin Compostion | 10-µm diameter nonporous polystyrene divinylbenzene substrate (55% crosslinking) with poly(IDA) grafts. | | Metal Ion Capacity (µmol Cu) | > 80 mmol Cu (4 x 250 mm analytical column) | | Chelating Metals | Ni, Fe, Cu, others | | Protein Loading Capacity (mg/mL gel) | >60 mg Lysozyme | | Dimensions | ProPac IMAC-10: 1 x 50 mm, 2 x 50 mm, 4 x 50 mm, 9 x 50 mm ProPac IMAC-10: 4 x 250 mm, 9 x 250 mm, 22 x 250 mm |
| | Order Information | | ProPac IMAC-10 Column (2 x 50 mm) | 063272 | | ProPac IMAC-10 Column (4 x 50 mm) | 063276 | | ProPac IMAC-10 Column (9 x 50 mm) | 063615 | | ProPac IMAC-10 Column (4 x 250 mm) | 063278 | | ProPac IMAC-10 Column (9 x 250 mm) | 063280 | | ProPac IMAC-10 Column (22 x 250 mm) | 063282 | | ProPac IMAC-10 Column (1 x 50 mm) | 063617 |
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