Science Catalog 2012 Life Cell Signaling Protein Labeling and Detection HaloTag® Fluorescent Ligands Product HaloTag® TMR Ligand HaloTag® Oregon Green® Ligand HaloTag® diAcFAM Ligand HaloTag® Coumarin Ligand HaloTag® Alexa Fluor® 488 Ligand HaloTag® TMRDirect™ Ligand HaloTag® R110Direct™ Ligand HaloTag® Biotin Ligand HaloTag® PEG-Biotin Ligand Size Conc. Cat.# Price (Fr) 30 µl 5 mM G8251 831.00 15 µl 5 mM G8252 437.00 30 µl 1 mM G2801 831.00 15 µl 1 mM G2802 437.00 30 µl 1 mM G8272 831.00 15 µl 1 mM G8273 437.00 30 µl 10 mM G8581 831.00 15 µl 10 mM G8582 437.00 30 µl 1 mM G1001 831.00 15 µl 1 mM G1002 437.00 30 µl 0.1 mM G2991 489.00 30 µl 0.1 mM G3221 489.00 30 µl 5 mM G8281 831.00 15 µl 5 mM G8282 437.00 30 µl 5 mM G8591 831.00 15 µl 5 mM G8592 437.00 For Research Use Only. Not for Use in Diagnostic Procedures. Description: The HaloTag® Fluorescent Ligands can carry a variety of functionalities, including fluorescent labels, affinity tags and attachments to a solid phase. The covalent bond forms rapidly under general physiological conditions, is highly specific and essentially irreversible. The HaloTag® Fluorescent Ligands allow researchers to apply the chloroalkane group that HaloTag® protein reacts with to any compound or surface with a compatible chemical group, creating endless possible applications. HaloTag® Fluorescent Ligands for Cellular Imaging Cell-permeant fluorescent ligands (rapid labeling protocol): • HaloTag® TMR Ligand (555Ex/585Em) • HaloTag® Oregon Green® Ligand (496Ex/516Em) • HaloTag® diAcFAM Ligand (494Ex/526Em) • HaloTag® Coumarin Ligand (353Ex/434Em) Cell-impermeant fluorescent ligands for cell-surface labeling (rapid labeling protocol): • HaloTag® Alexa Fluor® 488 Ligand (494Ex/517Em) Cell-permeant fluorescent ligands (“no wash” protocol): • HaloTag® TMRDirect™ Ligand (555Ex/585Em) • HaloTag® R110Direct™ Ligand (502Ex/527Em) The Alexa Fluor® 488 Ligand is impermeable to cell membranes and, therefore, used to label cell surface proteins. The TMR Ligand, Oregon Green® Ligand, diAcFAM Ligand and Coumarin Ligand readily cross the cell membrane and, therefore, can be used to label intracellular proteins. HaloTag® Ligands for Protein Detection The HaloTag® Biotin Ligand consists of a 12-atom linker arm to biotin and is used as an affinity tag to capture the HaloTag® protein-based fusion construct using the strong biotin-streptavidin interaction. The HaloTag® PEG-Biotin Ligand contains a spacer not found in the HaloTag® Biotin Ligand. This provides a significantly longer and more flexible linker between streptavidin and the HaloTag® protein, which may be advantageous in preserving the activity of a HaloTag® fusion partner protein upon immobilization or derivatization. Colabeling of HaloTag®-p65 fusion protein with HaloTag® TMR Ligand and the Anti-HaloTag® pAb. Panel A. Cytoplasmic (red) labeling of HEK293-p65-HT2 cells by HaloTag® TMR Ligand. Panel B. Cytoplasmic (green) labeling by Anti-HaloTag® pAb and Alexa Fluor® 488-conjugated antirabbit IgG (Invitrogen). Panel C. Colocalization of ligand and antibody binding activities. Panel D. Merger of red and green fluorescence with counterstaining of the nucleus by DAPI (blue). Arrows denote rare cells that show little or no expression of HaloTag®-p65. Protocols developed and performed at Promega. C. D. A. B. Features: • Label in Solution or on a Solid Support: The HaloTag® Ligands bind to the HaloTag® protein or protein fusions with high specificity and affinity. • Label Your HaloTag® Protein in Live Cells: The HaloTag® TMR, diAcFAM, Coumarin and Biotin Ligands readily cross the cell membrane. • Pull Down Protein Complexes: The spacer and reactive linker of the HaloTag® PEG-Biotin Ligand provide ideal pull-down capabilities. Alternatively, pull down directly with the HaloLink™ Resin. • Image Fixed Cells: The covalent bond is stable, allowing imaging of fixed cells and analysis of the labeled protein under stringent conditions. • Introduce Novel Functionalities or Perform Sequential Labeling: The open architecture of the technology enables the use of different ligands for multiple applications. • Design Only One Genetic Construct for Multiple Experiments: Obtain new functionality by using a different HaloTag® Ligand without having to design and clone a new expression construct. Protocol HaloTag® Technology: Focus on Imaging Technical Manual HaloTag® Biotin Ligand Protocol HaloTag® PEG-Biotin Ligand Protocol Part# TM260 9PIG828 9PIG859 236 For complete and up-to-date product information visit: www.promega.com/catalog 6312TB Pagina 239
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