Creative Biolabs is offering the most comprehensive services for antibody development projects. With strict regulation and effective execution, we are dedicated to providing the most valuable solutions to complete your projects.
HighlightsWith over a decade of experience in phage display technology, Creative Biolabs can provide a series of antibody or peptide libraries that are available for licensing or direct screening. These ready-to-use libraries are invaluable resources for isolating target-specific binders for various research, diagnostic or therapeutic applications. Highlights
Creative Biolabs has established a broad range of platforms for developing novel antibodies or equivalents. These cutting-edge technologies enable our scientists to meet your demands from different aspects and tailor the most appropriate solution that contributes to the success of your projects.
HighlightsWith deep understanding in antibody-related realms and extensive project experience, Creative Biolabs offers a variety of references to help you learn more about our capacities and achievements, including infographic, flyer, case study, peer-reviewed publications, and all kinds of knowledge that can assist your projects. You are also welcome to contact us directly for more specific solutions.
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HighlightsRecognized as a reputable protein-protein interaction assay service provider, Creative Biolabs is committed to offering comprehensive yeast two-hybrid (Y2H) solutions, one of the most standardized in vivo approaches for protein-protein and interaction screening and validation, for the discovery or validation of new interactions. Both the GAL4 Y2H system for nucleoprotein interaction screening and the DUAL Y2H system for membrane protein interaction screening are available based on our optimized Y2H platform.
All processes in living cells virtually depend on protein-protein interactions (PPIs). Thus, molecular biology and disease research need to understand PPI networks. Yeast two-hybrid (Y2H) is an extremely powerful technique for studying protein-protein interactions and especially, can be used to search for a novel interacting partner by screening a single protein or domain against a library of other proteins. It is this latter characteristic that finding the interacting protein without any prior identity of such proteins is the strongest application of Y2H assay.
Y2H as a cogent genetic system is one of the most standardized approaches currently for mapping PPIs both at a small scale and in a high throughput manner. In Y2H screening, PPIs are detected by the activation of reporter genes which respond to a reconstituted transcription factor (TF). Y2H is commonly applied to test some "prey" proteins for interactions with a single "bait" or target protein or pool of proteins. The merit of this method is the direct identification of interacting protein pairs without downstream experiments since the prey protein has been known and does not need further confirmation.
Fig.1 A simple flowchart for a random yeast two-hybrid (Y2H) library screening service. (Creative Biolabs)
Traditional Y2H system for PPI detection was developed based on the principle that the Gal4 transcription factor in the yeast Saccharomyces cerevisiae was composed of two functional domains, one is the DNA-binding domain (BD) mediating and the other is the activation domain (AD) responsible for transcription activation. DNA-BD located in the N-terminal 1-147 amino acid residues can recognize and bind to the upstream activating sequence (UAS) of the GAL4 response gene. AD located in the C-terminal 768-881 amino acid residues is responsible for initiating the downstream gene transcription of UAS by binding to other components in the transcriptionmachinery. Either DNA-BD or AD can't activate the transcription when functions separately. Only the two domains are sufficiently close in space, will they represent complete GAL4 transcription factor activity and activate the UAS downstream promoter, so that the downstream gene of the promoter can be transcribed.
Fig.2 The yeast two-hybrid system is a genetic tool used to detect interactions between two proteins. (Mehla, 2015)
Due to the limitations of the traditional GAL4 Y2H system, which is only available for the PPI detection of nucleoproteins, the novel DUAL Y2H technology for the PPI detection of membrane proteins has been developed, expanding the application range of Y2H. Different from the GAL4 Y2H system based on the Gal4 transcription factor, the DUAL Y2H system screens the PPI based on the principle of split-ubiquitin. The wild-type ubiquitin is a highly conserved protein with Nub terminus and Cub terminus, playing important roles in the degradation of proteins. To apply for the PPI detection, the isoleucine at position 3 of ubiquitin Nub was mutated to glycine (NubI was mutated to NubG), leading to a significant reduction in the affinity and no reassociation between Nub and Cub, thereby resulting in abnormal function of ubiquitin.
Generally, the bait protein of interest and prey library/protein are fused to the Cub and NubG respectively. The NubG and Cub will be brought together into proximity once the bait and prey interact with each other, thus resulting in the reconstitution of a full-length and normal function of ubiquitin. The reconstituted ubiquitin is recognized and catalyzed by ubiquitin-specific proteases (UBPs), releasing the transcription factor, which further enters the nucleus and activates the transcription of reporter genes.
Fig.3 Main components of the membrane-based split-ubiquitin Y2H system. (Ivanusic, 2015)
Creative Biolabs has completed plenty of protein-protein interaction screening or validation based on yeast hybrid systems, accumulating abundant expertise and experience in protein-protein interaction identifications. Added by rich experience and the perfect technology platform, our seasoned scientists offer one-stop or tailored Y2H services on a strictly fee-for-service basis. Both the GAL4 Y2H system and the DUAL Y2H system are available based on your needs.
Creative Biolabs optimized Y2H protocols. The efficiency and quality of our Y2H system have been satisfying clients with highly reproducible and reliable results. We are also skilled at other types of two-hybrid programs, such as mammalian two-hybrid and bacterial two-hybrid services, to suit worldwide scientists' needs in a wide range of biological fields. Please contact us for more details.
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All listed services and products are For Research Use Only. Do Not use in any diagnostic or therapeutic applications.
