HaloTag® Complete Pull-Down System Technical Manual
Literature # TM360
The HaloTag® Complete Pull-Down System provides the necessary reagents for the isolation and identification of intracellular protein complexes from mammalian cells. A protein-coding region of interest is cloned initially into the amino (N)-terminal pFN21A HaloTag® CMV Flexi® Vector or carboxy (C)-terminal pFC14K HaloTag® CMV Flexi® Vector or both for expression of HaloTag® fusion proteins under a CMV promoter when transfected into mammalian cells. The expressed HaloTag® fusion protein and the endogenous interacting protein partners are isolated as a protein complex. The HaloTag® fusion protein forms a highly specific and irreversible bond with the HaloLink™ Resin, allowing rapid and efficient capture of dilute protein complexes. As a negative control for the pull-down assay, HaloTag® Control Vector is provided. To more fully characterize the function of the protein of interest, the HaloTag® TMRDirect™ Ligand allows the study of cellular localization, trafficking and protein turnover using the same HaloTag® construct.