Cell Signaling MagneHis™ Protein Purification System Product MagneHis™ Protein Purification System Available Separately MagneHis™ Ni-Particles For Research Use Only. Not for Use in Diagnostic Procedures. Description: The MagneHis™ Protein Purification System provides a simple, rapid and reliable method for the purification of polyhistidine- or HQ-tagged, expressed proteins. Paramagnetic precharged nickel particles (MagneHis™ Ni-Particles) are used to isolate polyhistidine- or HQ-tagged protein directly from a crude cell lysate using either a manual (requires use of a magnetic stand) or automated procedure. Using a tube format, polyhistidineor HQ-tagged protein can be purified on a small scale using less than 1ml of culture or on a large scale using more than 1 liter of culture. Samples can be processed in a high-throughput manner using a robotic platform such as the Beckman Coulter Biomek® 2000 or FX. Features: • Simple: No centrifugation or vacuum is required once the cells are lysed. • Flexible: MagneHis™ Ni-Particles are compatible with a variety of common buffers. • Quick: No long incubations with lysozyme are required for cell lysis. • Scalable: Volumes can be adjusted to correspond to the amount of material to be purified, 1ml to 1 liter of cell culture. • Efficient: Binding capacity is up to 1mg of polyhistidine-tagged protein per 1ml of MagneHis™ Ni-Particles. • Versatile: Purification can be performed manually (requires use of a magnetic stand) or using an automated platform. • Convenient: Complete system that includes all necessary components, including a unique lysis buffer. • Automate This Assay: Validated automated methods are available for Beckman Coulter Biomek® 2000 and FX and Tecan Genesis® RSP at: www.promega.com/automethods/ Storage Conditions: Store at 4°C. Protocol MagneHis™ Protein Purification System Technical Manual MagZ™ Protein Purification System Product MagZ™ Protein Purification System For Research Use Only. Not for Use in Diagnostic Procedures. Description: The MagZ™ Protein Purification System provides a simple, rapid and reliable method for the purification of expressed polyhistidine- or HQ-tagged proteins, which are 99% free of hemoglobin contamination, from rabbit reticulocyte lysate. Based on the use of proprietary, paramagnetic precharged particles, polyhistidine- or HQ-tagged protein can be isolated from 50–500μl of TNT® Coupled Transcription/Translation reactions. Polyhistidine- or HQ-tagged proteins bind to the particles in minutes, while unbound proteins are washed away, and the target protein is eluted with imidazole. Features: • Specific: Minimal hemoglobin (<0.1%) binding to the MagZ™ Binding Particles. • Quick: No long incubations are required. • Versatile: Binding/wash and elution conditions can be further optimized for individual polyhistidine- or HQ-tagged proteins. Storage Conditions: Store at 4°C. Protocol MagZ™ Protein Purification System Technical Bulletin Part# TB336 Size Cat.# Price (Fr) 30 reactions V8830 433.00 Part# TM060 Size Cat.# Price (Fr) 65 reactions V8500 339.00 325 reactions V8550 1441.00 2 ml V8560 248.00 10 ml V8565 1054.00 HisLink™ Protein Purification Resin Product HisLink™ Protein Purification Resin For Research Use Only. Not for Use in Diagnostic Procedures. Description: HisLink™ Protein Purification Resin is a macroporous silica resin modified to contain a high level of tetradentate chelated nickel (>20mmol Ni/ml settled resin). The resin is designed to efficiently capture and purify bacterially expressed polyhistidine- or HQ-tagged proteins. The HisLink™ Resin also may be used for general applications requiring an immobilized metal affinity chromatography (IMAC) matrix. HisLink™ Resin may be used in either column or batch purification formats. The lysate flows by gravity over a column of packed HisLink™ Resin at a rate sufficient for complete capture and efficient elution of polyhistidine- or HQ-tagged proteins from cleared lysate. The resin may be used in vacuum filtration devices (e.g., Vac-Man® Vacuum Manifold [Cat.# A7231]), allowing simultaneous processing of multiple columns. HisLink™ Resin is also an excellent choice for affinity purification of polyhistidine- or HQ-tagged proteins with low- to medium-pressure liquid chromatography systems such as FPLC. In batch format, HisLink™ Resin may be separated easily from the lysate without filtration (or centrifugation), allowing processing of larger quantities of lysate and the ability to purify protein without clearing the lysate of insoluble cellular debris. Features: • Optimize Yields: High binding capacity (>15mg/ml). • Save Time: Purify polyhistidine- or HQ-tagged proteins from cleared or crude cell lysates. • Flexible: Use standard gravity column chromatography or automated applications such as FPLC. Storage Conditions: Store at 4°C. Protocol HisLink™ Protein Purification Resin Technical Bulletin HisLink™ 96 Protein Purification System Product HisLink™ 96 Protein Purification System For Research Use Only. Not for Use in Diagnostic Procedures. Description: HisLink™ 96 Protein Purification System provides a simple, quick and robust method of purifying multiple polyhistidine- or HQ-tagged expressed proteins from E. coli using a vacuum-based method. The system is designed to purify expressed polyhistidine- or HQ-tagged proteins directly from deep-well, 96-well plates. The HisLink™ 96 System is amenable to manual or automated methods, such as the Beckman Coulter Biomek® 2000 or FX for high-throughput applications. Using the provided FastBreak™ Cell Lysis Reagent, bacterial cells containing polyhistidine- or HQ-tagged protein are lysed directly in culture. The HisLink™ Resin is added directly to the lysate, and the polyhistidine- or HQ-tagged proteins bind in a matter of minutes. Samples are then transferred to a filtration plate, unbound proteins are washed away and the target protein is recovered by elution with imidazole. Features: • Simple: No centrifugation required; lysis buffer is added directly to cells in culture medium. • Quick: No long lysozyme incubations are required for cell lysis. • Versatile: Perform purification manually or on an automated platform. • Efficient: Binding capacity of 1mg of polyhistidine-tagged protein per well. Storage Conditions: Store the system at 4°C. Plates may be stored at 4°C or room temperature. Protocol HisLink™ 96 Protein Purification System Technical Bulletin For complete and up-to-date product information visit: www.promega.com/catalog Part# TB342 253 Size Cat.# Price (Fr) 1 × 96 blank V3680 499.00 5 × 96 blank V3681 2121.00 Part# TB327 Size Cat.# Price (Fr) 5 ml V8823 69.00 50 ml V8821 621.00 14 Protein Purification and Interactions Pagina 256
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