Protein is the ultimate executor of cell activity and function. Each protein does not perform its function independently. They usually interact with other proteins in cells to form a large complex to complete specific functions in specific time and space. Therefore, studying the interactions between proteins is of great significance for gaining insight into many life processes. A research hot spot in the post-genome era is to explore protein-protein interactions and their roles in regulating various life activities.
Methods
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Co-Immunoprecipitation
Co-IP is a technology that precipitates the protein that interacts with the antibody target protein in the sample together with the immune complex, and then detects it. Co-immunoprecipitation can be used to detect whether two target proteins interact in vivo (directly or indirectly). After co-immunoprecipitation, mass spectrometry can also be used to find an interacting protein in vivo.
Fig. 1 The basic flowchart of the co-immunoprecipitation technique.
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GST pull down
Pull down technology uses immobilized, labeled bait protein or tag protein to catch interacting proteins from cell lysate. This technology can determine the interaction relationship between known proteins and caught proteins or purified related proteins, and detect protein interaction relationships from in vitro transmission or translation systems.
Fig. 2 The basic flowchart of the GST pull down technique.
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Yeast Two-hybrid
Yeast two-hybrid is a method for identifying protein-protein interactions using recombination of transcriptional activators in yeast. This method measures the ability of two proteins to interact, and a transcriptional reporter system can be used to evaluate this interaction. In addition, this method can also be used to screen proteins with unknown interactions from protein libraries.
Fig. 3 The basic flowchart of the Yeast Two-hybrid technique.
Applications
- From protein interaction to biological function: using direct protein interaction, discovering interacting proteins, and then analyzing the biological function of these proteins
- From biological function to protein interaction: by changing biological phenomena, analyzing the genetic interaction between proteins, and further studying the interaction of related proteins
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Water Bath
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Centrifuge
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Autoclave Sterilizer
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Ultra-Clean Bench
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Laboratory Incubator
An incubator is a device used to grow and maintain microbiological cultures or cell cultures. It is made up of a chamber with a regulated temperature. An incubator maintains optimal temperature, humidity, and other conditions such as the CO2 and oxygen content of the atmosphere inside. Incubators are essential for much experimental work in cell biology, microbiology, and molecular biology.
STEMart provides you with a variety of protein-protein interaction technology equipment or consumables to meet your various R&D and application needs. If you have any questions or requirements for molecular interactions technology, please feel free to contact us.