Nanjing Xinfan Bio: Monitoring Protein Interactions at Live Cell Levels |

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Traditional methods for studying protein interactions are usually performed in vitro, using only protein fragments and not providing data in the context of the cellular environment. However, the lack of test results at the cellular level is not convincing, and cytological tests are needed to verify. For NanoBRETTM protein interaction analysis, researchers were able to study the induction and inhibition of protein interactions at the living cell level using full-length proteins.

The principle of BRET is: when two protein molecules interact, the luminescence signal spectrum generated by the enzymatic reaction of a protein-fused donor luciferase protein (traditional using Renilla Renilla luciferase) and another protein marker The excitation spectra of the acceptor fluorophores overlap, inducing the acceptor molecule to fluoresce. Compared to fluorescence resonance energy transfer (FRET), BRET does not require excitation light and has a lower background.

NanoBRETTM analysis is a greatly improved BRET assay platform using NanoLuc® luciferase as an energy donor and the HaloTag protein-labeled NanoBRETTM 618 fluorophore as a receptor. When the bright blue-shifted luminescent signal from the NanoLuc donor is coupled to the far-shifted HaloTag receptor, the spectral superposition is better, the signal is stronger, and the background is lower.

Compared to the traditional BRET method, the patented NanoLuc® luciferase protein as a donor in the NanoBRETTM assay is more optically active and ideally matched to the wavelength of the acceptor fluorophore. It uses the advanced Flexi vector technology for cloning, which is simpler and less time consuming. At the same time, Promega provides a commercial pre-built carrier, which saves time for self-construction of the carrier and is more convenient for application.

The Flexi Vector Cloning System is a restriction enzyme based on two rare restriction sites, SgfI and PmeI, which transfer the protein coding region between different Flexi. vectors quickly, efficiently and with high fidelity without resequencing . The NanoBRETTM PPI Flexi Starter System provides a solution for protein mutual detection based on the Flexi vector system, including vectors, detection reagents and positive controls.

Another solution for building your own carrier is the NanoBRETTM PPI MCS Starter System. This kit provides a NanoLuc® fusion protein cloning vector with a multiple cloning site and a HaloTag® fusion protein vector, detection reagents and a positive control.

Of course, you can also choose to pre-build the carrier. The NanoBRETTM pre-constructed vector fuses the genes of known interacting proteins. You don't need to rebuild the vector by yourself. It can be directly used to construct a cell model for transfection, and the mechanism of action and other related mechanisms can be studied with the detection reagents. Save time and effort, and the results are more guaranteed.

Elisa kit, imported elisa kit, elisakit, imported serum, imported reagent, gelatin zymography test kit, rat elisa test kit, human elisa test kit-Nanjing Xinfan Biotechnology Co., Ltd.