Our Protein Interaction Services use a affinity tag based purification technology platform specially designed for high throughput analysis and identification of protein: protein & protein:rna interactions.
Defining protein & ribonucleoprotein complexes is critical to virtually all aspects of cell biology, because many cellular processes are regulated by stable protein complexes and their identification often provides insights into their function. Our technology platform can also be used to study disease relevant protein & ribonucleoprotein complexes in order to fully understand the molecular mechanisms of diseases.
Our protein interaction service involves the characterization of protein & ribonucleoprotein complexes by expression of bait-proteins in mammalian cells. These baits are tools to isolate cellular proteins and their natural interactions partners as protein or ribonucleoprotein complex. They are isolated through affinity tag purification methods and subsequent identification of interaction partners through mass spectrometry, immunoprecipitation and Western blot analysis. After the isolation and identification of protein complexes, a functional analysis using RNA interference (RNAi) allow us to obtain more information about the cellular function of the protein complexes.
Interactome Inhibition
Innoprot protein interaction service includes the development of specific protein interactions assays; which can be customized and screened using our broad range of cell lines & primary cells.
A functional analysis using RNA interference allows to obtain more information about the cellular function of the protein complexes. We can inhibit the expression of specific protein/s performing related assays; and then study the behavior of the knockdown cells. If necessary, we can also provide you with the stable knockdown cell line.
Interactome Validation
Once the interacting proteins have been identified, we offer the possibility of checking the association of some specific interacting partners in the complexes, following the next steps:
- Immunoprecipitation of the “bait protein” using specific antibodies against new identified partners
- Western blot analysis
Interactome Identification
Our Interactome Identification service allows the identification of interacting proteins in a native cellular environment through an “in vivo” purified system that includes the next steps:
- Tag Affinity Subcloning
- Expression of bait-proteins in mammalian cells
- Protein Complexes Isolation through affinity tag purification methods
- Identification of interaction partners through mass spectrometry
Optionally, we can also identify interacting RNAs by RT-PCR & resultant cDNAs Sequencing.
We can also generate a stable constitutive expression cell line that you can you use like endless and reliable material source in future experiments; as well as, to prevent the transfection problems.