Status:
ReviewerMember since: 2016
Organization: Sanford Burnham Prebys Medical Discovery Institute
Great results. I've used this for so many different projects.
Application Area: Protein interaction
"Excellent versatility and ease of use. Easy to make a toolbox of ppi sensors."
Status:
ReviewerMember since: 2019
Organization: NIH
Very sensitive detection method in live cells.
Application Area:Analyze protein-protein interaction
"This kit is to detect protein-protein interaction using luciferase activity. It is easy to make DNA constructs for fusion proteins. After a simple transfection of 2 DNA plasmids into mammalian cells, one can detect the interaction between 2 fusion proteins by monitoring luciferase activity. It is very sensitive and easy to use. When protein association is induced by some treatments, one can monitor this association in a real time. When I had difficulty in setting up the system, Promega’s customer service helped me a lot."
Status:
ReviewerMember since: 2019
Organization: University of Cambridge
NanoBiT is an excellent system for studing PPI in real-time in live cells
Application Area:Monitor cell signalling protein-protein interaction in live cells in real-time
"NanoBiT system allows flexible development of a very effective and easy to use real-time protein:protein interaction assay. The starter kit contains MCS vectors for flexible cloning of any gene of interest with either N- or C-terminally positioned tags - fragments of NanoLuc. Upon interaction of two proteins of interest, fragments of split NanoLuciferase complement forming a fully active enzyme. NanoLuc activity is quantified with a specific substrate, furimazine. Luminescence detected with NanoLuc (used in NanoBiT) system is much stronger than of any other "classical" luciferase. This allows for the detection of protein:protein interaction which may often be rather transient and subtle cellular events. Also, it enables the expression of the interacting proteins at relatively low levels, as overexpression may lead to "forced" interaction and in result generate false positives. We developed multiple assays based on NanoBiT system for studding GPCR biology, including beta-arrestin recrutment and G-protein interaction assays among others."
Status:
ReviewerMember since: 2016
Organization: Vertex Pharmaceuticals
Great product; I would like to get more information on the use of this system for compound screening
Application Area:Protein-protein interaction in cells
"Quality is high with reproducible results. Only issue I have is specificity with small molecules."
Status:
ReviewerMember since: 2013
Organization: Swisspharma Research Laboratories,Inc.
Highest quality products that give excellence in our laboratory
Application Area:analyze all clinical samples with accuracy and precision.
"High throughput in efficiency and effectiveness for patientcare "
Status:
ReviewerMember since: 2016
Organization: Vertex Pharmaceuticals
Ease of use; low amounts of plasmids for transfection.
Application Area:Protein-protein interaction
"Dynamic range is about 3 order of magnitude compared to the background signal. It is easy to use and uses nanogram amounts of plasmids for transfection. The protein-protein interaction can be evaluated in 1:1 stoichiometry. I would certainly recommend this product to my colleagues. There is a good product and technical support from Promega."
Status:
ReviewerMember since: 2016
Organization: Institute for In Vitro Sciences
Excellent product.
Application Area:Protein-protein interaction, such as fret assay.
"Very efficient, easy to use, easy to analyze data. I will definitely recommend it to my colleagues."
Status:
ReviewerMember since: 2016
Organization: Sanford Burnham Prebys Medical Discovery Institute
Very easy to clone multiple tag locations.
Application Area:Protein Protein interaction HTS
"Very easy to clone multiple tag locations. Easy to use reagents with high signal to background. Excellent support. The flexi system makes cloning of all the combinations for testing very easy. Also, they have both strong and weak promoter systems to minimize and tune the expression level."
Status:
ReviewerMember since: 2016
Organization: UMASS Medical School
Greatly improves luciferase complementation assays.
Application Area:Protein-protein interaction upon cell stimulation.
"The small size of the bits increases stability of your fusion proteins. It is better than the original firefly luciferase complementation assay."
Status:
ReviewerMember since: 2016
Organization: Ryerson University
Excellent product and great results.
Application Area:Detecting protein-protein interactions between two trans-membrane proteins.
"It's a very easy to use technique. Protocol is very clear and fast. Controls work great and the quality of the results obtained are fantastic. I highly recommend this product for the study of protein - protein interactions."
Status:
ReviewerMember since: 2016
Organization: China Agriculture University
Excellent product and great results.
Application Area:Detecting protein-protein interactions between two trans-membrane proteins.
"It's a very easy to use technique. Protocol is very clear and fast. Controls work great and the quality of the results obtained are fantastic. I highly recommend this product for the study of protein - protein interactions."
Status:
ReviewerMember since: 2016
Organization: Yale University
Easy to use.
Application Area:Ligand-Receptor Interaction
"With modern DNA assembly tools this system is easy to use and even applicable for HTS."
Status:
ReviewerMember since: 2016
Organization: Cincinnati Childrens
Wonderful product. Easy to use.
Application Area:Protein interaction.
"It works well and it is super-bright compared to FRET."
NanoLuc® Binary Technology (NanoBiT) is a two-subunit system based on NanoLuc® luciferase that can be applied to the intracellular detection of protein:protein interactions (PPIs) in live cells. The NanoBiT® system is composed of two small subunits, Large BiT (LgBiT; 18kDa) and Small BiT (SmBiT; 11 amino acid peptide), that are expressed as fusions to target proteins of interest. The LgBiT and SmBiT subunits have been independently optimized for stability and minimal self-association. Interaction of the target proteins facilitates subunit complementation to give a bright, luminescent enzyme.
The NanoBiT® PPI Starter Systems provide the vectors required to create the LgBiT and SmBiT protein fusions, a PRKACA:PRKAR2A constitutively interacting positive control pair and a negative control vector. Starter systems also include the Nano-Glo® Live Cell Assay System, a single-addition, non-lytic detection reagent used for monitoring NanoBiT® luminescence in living cells. The reagent is prepared by diluting the Nano-Glo® Live Cell Substrate with the Nano-Glo® LCS Dilution Buffer to make the Nano-Glo® Live Cell Reagent. Both substrate and buffer solutions are optimized to provide enhanced stability and reduce autoluminescence in the presence or absence of serum, increasing the sensitivity for detection of low levels of NanoBiT® luminescence. The FKBP:FRB pair is provided separately as an inducible positive control.
Expression is driven by HSV-TK promoter, providing constitutive, low-level expression in mammalian cells. Using the NanoBiT® MCS Starter System, you can generate N- and C-terminal LgBiT and SmBiT fusions to proteins of interest using traditional cloning with a multiple cloning site (MCS). Using the NanoBiT® Flexi® Starter System, you can generate N- and C-terminal LgBiT and SmBiT fusions using the Flexi® Vector Cloning System, a directional cloning method based on two rare-cutting restriction enzymes, SgfI and PmeI, that provides a rapid, efficient and high-fidelity way to transfer protein-coding regions between Flexi® Vectors without the need to resequence. Utilize Find My Gene™ to obtain your ORF clones already in Flexi® format for simple creation of fusions.
Features:
Applications: