Cell Signaling HaloLink™ Protein Array Systems Product HaloLink™ Array (TNT® T7 Quick) Two Slide System HaloLink™ Array (TNT® SP6 Wheat Germ) Two Slide System HaloLink™ Array Six Slide System HaloTag® Standard Protein For Research Use Only. Not for Use in Diagnostic Procedures. Description: Protein arrays enable parallel analysis of multiple protein:protein, protein:drug or protein:nucleic acid interactions. The HaloLink™ Protein Array Systems provide a new way to create homebrew (on-demand) protein arrays by combining innovative HaloTag® technology, surface engineering and cell-free protein expression systems. The HaloTag® protein is a mutated hydrolase that forms a covalent bond with HaloTag® ligands. Under physiological conditions binding is rapid and highly specific, yielding a complex that is stable even under stringent conditions. Using the HaloLink™ Protein Array Systems, HaloTag® fusion proteins are expressed in a cell-free expression system and then covalently captured on hydrogelcoated glass slides derivatized with HaloTag® Ligands. The fusion proteins are captured directly from the expression reaction mixture without prior purification. Using this approach, multiple fusion proteins can be rapidly synthesized and immobilized in parallel on the slide surface, and an entire experiment including protein expression, custom array formation and protein interaction analysis can be completed in less than eight hours. The HaloLink™ Array Two Slide Systems (Cat.# G6140 and G6180) contain HaloLink™ Slides, HaloLink™ Array Gaskets for creating 50-well arrays, a HaloTag® Standard Protein, Anti-HaloTag® Antibody and a cell-free expression system [either the TNT® T7 Quick Coupled Transcription/Translation System (Cat.# G6140) or the TNT® SP6 High-Yield Wheat Germ Protein Expression System (Cat.# G6180)]. The HaloLink™ Array Six Slide System contains HaloLink™ Slides, HaloLink™ Array Gaskets and Anti-HaloTag® Antibody. To use the Six Slide System you will need to provide your own protein expression system or order the TNT® T7 Quick Coupled Transcription/Translation System (Cat.# L1170 or L1171) or TNT® SP6 High-Yield Wheat Germ Protein Expression System (Cat.# L3260 or L3261). The HaloTag® Standard Protein (Cat.# G4491) is not included with the Six Slide System but can be ordered separately. The HaloTag® technology offers a quick and convenient way to test protein expression of HaloTag® fusion proteins by labeling with fluorescent HaloTag® TMR Ligand followed by SDS-PAGE analysis. Guidelines can be found in the HaloLink™ Protein Array Technical Manual #TM310. Size Cat.# Price (Fr) 2 arrays G6140 638.00 2 arrays G6180 638.00 6 slides G6190 1245.00 30 µg G4491 92.00 Features: • Fast Protein Production: Cell-free expression systems allow quick, single-tube, coupled transcription/translation for the production of the proteins of interest to be used in the protein array experiment. • Irreversible Binding of the Captured Protein: Unlike other affinity tags, which tend to dissociate from the surface, HaloTag® fusion proteins are covalently bound to the HaloLink™ Slide. • No Protein Purification Step: The protein of interest is immobilized directly from the cell-free expression reaction. • Reduced Nonspecific Binding: HaloLink™ Slides have a unique hydrogel coating that is designed to prevent nonspecific binding while preserving the functionality of specifically captured proteins. • Extensive Washing Allowed: Covalent binding of HaloTag® fusion proteins to the HaloLink™ Slide allows extensive, stringent washing that results in reduced background and a lower incidence of false positives. • No Need for a Robotic Arrayer: The unique 50-well configuration allows multiple interactions to be studied in parallel without the need for a complex robotic arrayer. Storage Conditions: Store the TNT® lysates at –70°C. Store the HaloTag® Standard Protein and the Anti-HaloTag® Antibody at –20°C. The HaloLink™ Protein Array Slides should be stored at –20°C and opened just before use. After opening, unused slides should be stored at –20°C and used within one month. Store the HaloLink™ Array Gaskets at room temperature. • Do not store the TNT® lysate at any temperature other than –70°C. Storage at other temperatures (e.g., –20°C) for even a short time will dramatically reduce activity. • Do not freeze-thaw the TNT® lysate more than two times. • Do not store the TNT® lysate in the presence of dry ice. Prolonged exposure to dry ice can cause significant loss of activity. Protocol HaloLink™ Protein Array Systems Technical Manual Part# TM310 14 For complete and up-to-date product information visit: www.promega.com/catalog 249 Protein Purification and Interactions Pagina 252
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