Efficient Isolation and Identification of Intracellular Protein Complexes from Mammalian Cells Using HaloTag® Technology
Part # PS098
Jacqui Mendez, Nancy Murphy, Danette D. Hartzell, Natasha Karassina, Geogyi Los, Marjeta Urh and Keith Wood
Promega Corporation, Madison, WI
- HaloTag (HT) fusion proteins form a highly specific and covalent bond with the HaloLink resin, allowing rapid capture of dilute protein complexes from cellular lysates in a single step purification method.
- Captured protein partners can be eluted using SDS or cleaved from the resin using TEV protease and analyzed by mass spectrometry for the identification of unknown binding partners or by Western blotting for the confirmation of known or suspected protein partners.
- The HaloTag technology is also applicable for the study of in vivo Protein:DNA interactions and cellular localization of the protein of interest using fluorescent HaloTag ligands.
- The ability to perform these different experiments with the same fusion protein eliminates the need to make multiple constructs for each desired study allowing flexibility to easily expand to other areas of research.
Printed in USA.