Halo tag TMR 

Halo tag TMR is a red fluorescent dye composed of Halo tag ligand molecules and TMR (TAMRA). Halo tag can quickly and stably covalently bind to Halo protein with high specificity and affinity (Ex/Em = 550/576 nm)^[1].

Guide (The following is our recommended protocol. This protocol is only a guide and should be modified according to your specific needs).
Staining steps^[1]:
1) Transfect cells with a plasmid encoding the target HaloTag fusion protein and culture to the desired confluence under appropriate conditions.
2) Replace 1/5 of the serum-free culture medium or PBS with 5× HaloTag TMR working solution to a final concentration of 5 μM. Mix by gently shaking the culture container.
3) Incubate the cells in a 37°C, CO₂ incubator for 15 minutes to allow the ligand to covalently bind to the HaloTag protein.
4) For permeable ligands: Remove the ligand-containing medium and replace with an equal volume of prewarmed serum-free culture medium or PBS. Repeat this wash step two more times for a total of three washes. For cell surface markers (non-permeable ligands): Wash twice with serum-free culture medium or PBS to remove excess ligand from the extracellular environment.
5) Incubate the cells in complete medium at 37°C for an additional 30 minutes to ensure complete removal of unbound ligand.
6) Replace the medium with phenol red-free medium (if available) to reduce autofluorescence.
7) Transfer the cells to a container suitable for imaging and observe using a fluorescence microscope.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

636.18

C₃₅H₄₂ClN₃O₆

2764890-88-0

Solid

Purple to purplish red

C/[N+](C)=C1C=CC2=C(C3=CC(C(NCCOCCOCCCCCCCl)=O)=CC=C3C([O-])=O)C4=CC=C(N(C)C)C=C4OC2=C/1

Room temperature in continental US; may vary elsewhere.

-20°C, protect from light, stored under nitrogen

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light, stored under nitrogen)

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light, stored under nitrogen). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

• [1]. Benink HA, et al. HaloTag technology for specific and covalent labeling of fusion proteins. Methods Mol Biol. 2015;1266:119-28.  [Content Brief]