signal transduction

Service details:
Cell signal transduction refers to the process that extracellular factors combine with receptors (membrane receptors or nuclear receptors) to trigger a series of biochemical reactions and protein-protein interactions in cells until the genes required for cell physiological reactions begin to express and various biological effects are formed It is known that there are many signal transduction modes and pathways in cells, and there are multiple levels of cross regulation between them. It is a very complex network system. Examples are as follows: The environment of higher organisms is changing all the time. The coordination and unity of body functions requires a perfect mechanism of mutual recognition, mutual reaction and interaction between cells. This mechanism can be called cell communication (cell communication).
In this system, cells or cells in contact are identified, Or recognize various signals in the surrounding environment (from surrounding or distant cells) and transform them into changes in various molecular functions in the cell, so as to change some metabolic processes in the cell, affect the growth rate of the cell, and even induce cell This change in the activity of various molecules against exogenous signals and the process of transmitting this change to effector molecules in turn to change cell function is called signal transduction (signal transduction). Its ultimate purpose is to make the body respond most appropriately to the changes of the external environment as a whole. In the regulation of material metabolism, it often involves the regulation of the metabolic pathway by the neuroendocrine system at the overall level.
Its essence is the process in which some cells send signals and others receive signals and transform them into changes in cell function. Therefore, to clarify the mechanism of cell signal transduction means to recognize the expression and regulation modes of cell proliferation, differentiation, metabolism and death in the whole life process, and then understand the regulation mechanism of body growth, development and metabolism. Detection of double luciferase reporter gene -- a sharp tool in the study of cell signal transduction Signal transduction detection Regulation (inhibition or enhancement) of cell signal transduction Regulation of target proteins on known proteins Promoter / enhancer detection Virus / cell interaction Transcription factors (interferon, NF-kB, IRF3) Drug induced action or effect Study on the regulation of cytokines by various radiation treatments Traditional detection method WB Necessary data for SCI articles (if > 3, regulation function research). Western blotting takes too long to get results The operation is cumbersome and requires the use of expensive antibodies WB results lack statistical basis. Too many human factors Advantages of luciferase reporter gene Superior sensitivity, more than 1000 times higher than the Western bath sensitivity. With reliable statistical principle, each group of experiments was repeated three times. High signal value, no endogenous activity. At present, the mainstream detection methods. Fast detection and high sensitivity. We do not give up WB detection. After luciferase detection, the sample is subject to WB detection, which makes the experiment more convincing. It has a wide range of applications and can detect any molecule in cells. Detection principle of luciferase reporter gene Principle: prepare DNA vector containing Luc / Rluc and overexpress the cell line. Cell selection: cell lines are selected according to experimental needs, usually 293T, Sheila cells and primary cells with high transfection efficiency. Transfection: the reporter gene and target gene labeled with luciferase were overexpressed (at different time points) Provide stimulation:
generally, it is stimulated by various conditions such as drug treatment, virus infection, UV irradiation and so on. Protein collection: the imported / domestic Kit (Promega, Invitrogen Shangbai Biology) was used for collection, and the experiment was repeated three times. Measuring fluorescence: modulus ™ Single tube multifunctional fluorometer for detection.
Service content:
1. We provide double fluorescein labeled reporter genes of IFN Luc, IFN Luc, ISRE Luc, prdii Luc, erse Luc, upre Luc and NF KB Luc- α、 IFN- β、 IRF3, NF-kB and other pathways were detected.
2. We provide common expression cells: 293T, Sheila and common primary cells.
3. Customers only need to provide target genes, fluorescein labeled reporter genes and expression cells according to experimental needs.
4. We provide real original data and result analysis report.
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