HaloTag® Ligands

Covalently Attach Fluorescent Labels or Affinity Tags to Membrane or Intracellular Proteins

• Cell permeant TMR, Oregon Green®, diAcFAM and Coumarin ligands readily cross the cell membrane for labeling intracellular proteins
• Cell impermeant tags include Alexa Fluor® 488 and 660 for quick cell surface labeling

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 (555_Ex/585_Em)
• HaloTag® Oregon Green® Ligand (496_Ex/516_Em)
• HaloTag® diAcFAM Ligand (494_Ex/526_Em)
• HaloTag® Coumarin Ligand (353_Ex/434_Em)

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

• HaloTag® TMRDirect™ Ligand (555_Ex/585_Em)
• HaloTag® R110Direct™ Ligand (502_Ex/527_Em)

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

• HaloTag® Alexa Fluor® 488 Ligand (494_Ex/517_Em)
• HaloTag® Alexa Fluor® 660 Ligand (663_Ex/690_Em)

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.

Applications

• 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.

temPart #Size

HaloTag® TMRDirect™ Ligand

G299A1 × 30μl

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