TRPV4-His499 protein sensitive to magnetic force, magnetic regulation tool and application
A technology of trpv4-his499 and magnetic regulation, applied in the field of system neuroscience, can solve the problems of consuming large magnetic nanoparticles and heat, specific modification of magnetic nanoparticles, and inability to accurately target cells
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[0033] In the present invention, the preparation method of the functionalized modified magnetic nanoparticles preferably includes: encapsulating the carboxylated magnetic nanoparticles with an anti-His antibody.
[0034] In the present invention, there is no special limitation on the source of the carboxylated magnetic nanoparticles, and commercial products can be used. In the present invention, there is no special limitation on the encapsulation method of the anti-His antibody on the carboxylated magnetic nanoparticles, and conventional methods can be used.
[0035] In the present invention, the anti-His antibody on the functionally modified magnetic nanoparticles can specifically react with the His tag of the TRPV4-his499 protein, so that the magnetic induction protein is fixed and dispersed on the cell surface, and under the action of a magnetic field, the The effector specifically opens TRPV4 protein ion channels, triggering cellular calcium influx for neural calcium signa...
Embodiment 1
[0042] 1. Preparation of experimental reagents
[0043] The carboxylated magnetic nanoparticles of 1.20nm, referred to as MNP (with ferric oxide core) were purchased from Chemicell, and were coated with antibodies according to the standard experimental method provided by Chemicell ( figure 1 shown), to obtain magnetic nanoparticles (MNPs). That is, under the catalysis of compounds such as EDC and NHS, the carboxyl groups on the magnetic nanoparticles and the amino groups on the His antibody can react to form amide bonds to achieve stable bonding and achieve antibody encapsulation of magnetic nanoparticles.
[0044] 2. The TRPV4-Hi499 fragment was synthesized by gene synthesis, that is, a 6×his tag was inserted at the 499th amino acid position of the TRPV4 protein, and the fragment was connected to the backbone plasmid through a restriction site to obtain pcDNA3.1-TRPV4- His499-p2A-mcherry, this plasmid is used for HEK293T cell transfection.
[0045] 3. Molecularly construct ...
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