Cell Signaling HaloTag® Mammalian Pull-Down Systems Product HaloTag® Mammalian Pull-Down and Labeling System HaloTag® Mammalian Pull-Down System HaloTag® Control Vector Available Separately Protease Inhibitor Cocktail, 50X Mammalian Lysis Buffer For Research Use Only. Not for Use in Diagnostic Procedures. Description: The HaloTag® Mammalian Pull-Down and Labeling System (Cat.# G6500 and G6504) is designed to capture and purify intracellular binary and higher order protein complexes, including transient or weakly interacting partners. This system includes the HaloTag® TMRDirect™ Ligand, which allows correlative cellular localization and real-time imaging studies with the same genetic construct as a means to further understand overall protein function. Cat.# G6504 does not include the TMRDirect™ Ligand. The HaloTag® Control Vector provides protein expression of the HaloTag® protein in mammalian cells, E. coli or in vitro expression systems dependent on human cytomegalovirus (CMV) intermediate early enhancer, T7 or SP6 RNA polymerase promoters. It can be used as a control for any HaloTag® experimental system and can be used for both stable and transient HaloTag® expression in mammalian cells; for stable expression, co-transfection with a vector containing a selectable marker is required. The Protease Inhibitor Cocktail, 50X, is a mixture of six different protease inhibitors with different target protease specificities. This product is provided in a freeze-dried format and can be reconstituted using either 100% ethanol or DMSO. For Expression-Validated HaloTag® ORF Clones visit Find My Gene at: www.promega.com/FMG fusion construct HaloTag® Transfection or stable expression Size Cat.# Price (Fr) 24 reactions G6500 490.00 24 reactions G6504 270.00 20 µg G6591 250.00 1 ml G6521 107.00 40 ml G9381 72.00 The Mammalian Lysis Buffer is designed for use with HaloTag® Mammalianbased expression systems such as the HaloTag® Mammalian Pull-Down and Labeling Systems (referenced here) as well as the HaloCHIP™ System (Cat.# G9410). Formulation consists of 50mM Tris-HCl, 150mM NaCl, 1% Triton® X-100 and 0.1% sodium deoxycholate (pH 7.5). Features: • Rapid, Efficient and Covalent Capture of Binary and Higher Order Complexes Directly from Lysates: Improved capture of protein partners, including transient interactions. • High Purity and Low Background: Improved accuracy in identification of proteins; covalent attachment allows bait protein to remain behind if desired. • Ability to Fluorescently Label the Same Genetic Fusion: Correlate complex capture with cellular localization. • Compatibility with All Downstream Methods of Analysis: Freedom to identify complexes in variety of applications including mass spectrometry. Storage Conditions: Store the 10X TBS Buffer and SDS Elution Buffer at room temperature. Store the HaloLink™ Resin and Mammalian Lysis Buffer at 4°C. Store the Protease Inhibitor Cocktail and HaloTag® TMRDirect™ Ligand at –20°C. Protocol HaloTag® Mammalian Pull-Down and Labeling Systems Technical Manual Part# TM342 14 HT POI and assemble protein complex. Express HaloTag® fusion protein HT POI Lyse cells. POI HT Protein of Interest HaloTag® Protein HT POI TEV cleavage POI SDS elution Elute with SDS buffer or cleave with TEV (optional). Analyze with downstream application. Representation of the HaloTag® mammalian pull-down assay using HaloTag® fusion protein as bait. HaloLink™ Resin Capture protein complex on HaloLink™ Resin. Wash HaloLink™ Resin. For complete and up-to-date product information visit: www.promega.com/catalog 247 Protein Purification and Interactions 9033MA POI HT Pagina 250
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