HaloTag Protein Reporter Cell Lines
Flexible and efficient …Wider experimental range…Reliable results
The combination of Horizon’s gene editing technology and Promega’s HaloTag platform enables researchers to study protein function at the endogenous level. There are two parts to the HaloTag technology: the HaloTag protein and the HaloTag ligands. The HaloTag ligands are designed to covalently bind the HaloTag protein and come in a variety of types including fluorescent ligands and solid substrates, enabling researchers to use HaloTag in a wide variety of applications such as live cell imaging and isolation of HaloTag and therefore the protein of interest.
HaloTag protein Reporter Cell Lines
In the HaloTag protein reporter cell lines, HaloTag has been introduced as either an N or C terminal fusion to the protein of interest at the native genetic locus.
In C-terminal fusions HaloTag is introduced directly upstream of the endogenous stop codon in the gene of interest, while in N-terminal fusions it is introduced directly downstream of the endogenous start codon. The configuration of the fusion is designed to ensure protein biology is not compromised, enabling the study of HaloTag-protein fusions at physiological levels in applications such as live cell imaging and isolation of proteins/protein complexes.
With HaloTag protein reporter cell lines you get:
- Temporal and spatial control of labelling enabled by the rapid labelling reaction, this results in precision in experiments interrogating timing of cellular events. Sequential labelling is also feasible in live cell imaging experiments.
- Labelling and imaging of live or fixed cells is permitted by all fluorescent ligands. The stability of the dyes and the HaloTag complex allows fixed cells to be imaged.
- Pull-down experiments are precise using the HaloTag Mammalian Pull-Down System and can identify intracellular complexes in the reporter cell lines. The complexes can be further analysed by a variety of methods including: western blotting, Mass spectrometry and SDS-PAGE.
Pull-down experiments with isogenic reporter lines demonstrate protein enrichment.
- HaloTag ligand linked to resin enables stringent pull-downs. High sensitivity via covalent interaction with matrix.
- Highly specific and sensitive analysis of HaloTag fusion proteins in western blots with a HaloTag specific antibody from Promega.
Live cell imaging of protein localisation (K-Ras) in the SW48 KRAS HaloTag protein reporter cell ine. Use of HaloTag ligand shows selective staining for HaloTag engineered lines over unmodified parental cells demonstrating specificity of signal. HaloTag ligand shows comparable results to ICC.
Control cell lines are also available where unfused HaloTag expression is driven by the promoter of the gene of interest. These are useful for demonstrating the observations made using HaloTag protein reporter cell lines are due to the intrinsic properties of the protein and not HaloTag itself.
For more information and/or to discuss experimental design: