Protein–protein interaction studies in living cells
Get expert help with your protein–protein interaction assays. Our protein-fragment complementation assay technology platform offers a broad range of solutions, including transfection-ready plasmids, stably transfected cell lines, and validated plasmid pairs.
Contact us to discuss the technology and options further. For more information, call 800-955-6288 x2 or email email@example.com. A project manager will respond to your inquiry within two business days.
Quantitative protein–protein interaction assays
Protein-fragment complementation assay (PCA) is the method of choice for the identification of protein–protein interactions in biological systems. Each target protein of interest is linked to one of two separate, inactive reporter subfragments. When the proteins of interest interact, the two fragments of the reporter come into close enough proximity to restore functionality to the reporter. The activity of the functioning reporter can be detected and quantified.
Since its initial discovery, PCA has been developed into a novel screening technology that has been deployed to investigate complex cellular biochemical networks, including disease-relevant pathways in living human cells. Additionally, the PCA technology has been used to screen for drugs and drug targets in the contexts of their native environments.
Choice of reporters
Dimerization-dependent Yellow Fluorescent Protein
Your target proteins are linked to two fragments of a Yellow Fluorescent Protein (Venus YFP). When the target proteins come into close proximity the two fragments of the Venus YFP form a functional reporter, and the interaction can be detected.
Dimerization-dependent luciferase reporter
Your target proteins are linked to either the N-terminal or C-terminal fragment of a luciferase reporter. When the target proteins come into close proximity the two fragments of the luciferase reporter form a functional reporter, and after the addition of the substrate the target protein–protein interaction can be measured and quantified.
Transfection-ready plasmids or stably expressing cell lines
- Plasmid constructions for transient transfection—the cDNAs expressing your target proteins of interest are cloned into plasmids that are ready for transfection and assay
- Stable cell line development—your target proteins and reporter of choice are stably expressed in your preferred cell line
Validated plasmid pairs are also available
In addition to custom cell line development, we also have several high-value protein–protein interaction targets already cloned into plasmids and ready for transient transfection assay development. These plasmid pairs have been developed and validated; if interested, contact us for a complete list of plasmid pairs available.
Contact us to discuss the technology and options further. A project manager will respond to your inquiry within two business days.
Phone (North America only): 800 955 6288 x2