The NaveniFlex in situ proximity ligation technology enables the visualization of protein-protein interactions and post-translational modifications through the use of two primary antibodies, one against each interacting protein, or against the protein and it’s modification. The technique can also be used to increase the sensitivity and specificity of your protein localization.
The Naveni Family
The NaveniFlex kits are based on a novel, proprietary proximity ligation assay principle which enables highly specific detection of proteins, protein interactions, or protein modifications in cultured cells as well as in fresh-frozen and FFPE samples.
The NaveniFlex kits provide accurate and reliable data with exceptionally precise staining patterns that can be quantified without image pre-processing. The NaveniFlex kits contain secondary detection reagents called Navenibodies, which are carefully selected antibodies conjugated to proprietary oligo arms. The Navenibodies can be used with one or two primary IgG antibodies raised in mouse (M), rabbit (R) or goat (G). Three fluorophore alternatives are included which allows you to choose the fluorophore most suitable to your microscope and assay. The working volume of the Navenibodies is 4000µl.
Visualize protein-protein interactions in the tissue microenvironment
The dual recognition approach enables visualization of protein-protein interaction through the use of one primary antibody for each target protein, without the need to disrupt the tissue microenvironment. See our application note on interacting glycoproteins in ovarian cancer. Other common methods for studying protein-protein interactions, like BRET, cross-linking, pull-down assays or co-immunoprecipitation (Co-IP) often require cell-modification or disruption of the tissue. This can lead to false positive results in interaction studies, since proteins that would not be in proximity in the intact tissue can be forced together in the lysate preparation or during cell processing.
Increase the specificity of your immunostaining with dual recognition
Correct localization of proteins is critical for both the understanding and treatment of many diseases. The quality of in situ protein detection assays depends mainly on the specificity of the antibodies used against the target. Many commercial antibodies have low specificity to the targets, generating unspecific background staining resulting in misleading results.
High specificity localization of post-translational modifications
This method also enables studies of post-translational modifications, for instance phosphorylation. The dual antibody approach allows you to combine a protein target specific antibody with a pan-specific phospho-antibody, and the proximity ligation technology results in specific detection of the phosphorylated protein.
What assay kit works with my primary antibodies?
If you have access to two primary antibodies, generated in rabbit, mouse or goat respectively, and want to study protein-protein interaction, increase the specificity and sensitivity of your protein detection, or investigate post-translational modifications like phosphorylation, the best choice is to use the NaveniFlex assay for the corresponding primary antibody pair. The NaveniFlex assays come in the following combinations: MR, GR and GM, containing Navenibodies against mouse and rabbit (MR), goat and rabbit (GR) and goat and mouse (GM) primary antibodies.
If you have access to just one primary antibody, and are looking to increase signal strength and signal to noise ratio, you can use the NaveniFlex MM or RR kits, with Navenibodies against mouse (MM) and rabbit (RR) primary antibodies.
See application notes and examples here
Custom assay development
If you have specific research questions, please contact us. We accommodate custom-made target specific assays and panel designs that can be set to suit your research project’s needs and requirements. Are you interested in our services? Get in touch to find out how we can help you.