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HaloTag TMRDirect Ligand

by Promega
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  • Overview
  • Protocols
  • Specifications
  • Resources
  • Ligands with a variety of functionalities, including fluorescent labels, affinity tags and solid phase attachment abilities. The ligands form covalent bonds rapidly under general physiological conditions; the bonds are highly specific and essentially irreversible. HaloTag® Fluorescent Ligands enable you to add the chloroalkane group with which HaloTag® protein reacts, to any compound or surface with a compatible chemical group. The possible applications are endless.

    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® Fluorescent Ligands for Cellular Imaging

    Cell-permeant 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-permeant ligands ("no wash" protocol):

    • HaloTag® TMRDirect™ Ligand (555Ex/585Em)

    • HaloTag® R110Direct™ Ligand (502Ex/527Em)

    Cell-impermeant ligands for cell-surface labeling (rapid labeling protocol):

    • HaloTag® Alexa Fluor® 488 Ligand (494Ex/517Em)
    • HaloTag® Alexa Fluor® 660 Ligand (663Ex/690Em)

    HaloTag® Ligands for Protein Detection

    • HaloTag® Biotin Ligand. Contains a 12-atom linker arm to biotin and is used as an affinity tag to capture the HaloTag® protein-based fusion construct via biotin-streptavidin interaction.
    • HaloTag® PEG-Biotin Ligand. Contains a spacer not found in the HaloTag® Biotin Ligand that provides a significantly longer, more flexible linker between streptavidin and the HaloTag® protein. This linker may help preserve HaloTag® fusion partner protein activity upon immobilization or derivatization.


    • 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.
    • Analyze Labeled Fusion Proteins Using SDS-PAGE, Mass Spectrometry, etc.: The bound ligand is stable under denaturing conditions.

    For more information, see the Protocols & Applications Guide.

    Structure of the HaloTag® Ligands showing the functional reporters and the reactive linker.
    Structure of the HaloTag® Ligands showing the functional reporters and the reactive linker.
  • Protocols

    Complete Protocol

    Download PDF

    HaloTag® Technology: Focus on Imaging Technical Manual

    PDF (1 MB)

    Download PDF

    HaloTag® Biotin Ligand Protocol

    PDF (128 KB)

    Download PDF

    HaloTag® PEG-Biotin Ligand Protocol

    PDF (128 KB)

    Video Protocols

  • Certificate of Analysis

    Lookup Certificate of Analysis

    Storage Conditions

    -30C TO -10C

    Use Restrictions

    For Research Use Only. Not for Use in Diagnostic Procedures.

  • Resources