Method for detecting if interaction between nucleic acid conjugated protein -target protein exist based on biological chip
A technology for binding proteins and target proteins, which is applied in the field of detecting whether there is an interaction between nucleic acid binding proteins and target proteins based on biochips, and can solve the problem that there is no high-throughput detection method for the interaction between nucleic acid binding proteins and target proteins. Achieve the effect of high sensitivity, strong specificity and broad application prospects
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Embodiment 1
[0038] Example 1. Detection of the specificity of the method for detecting whether there is an interaction between the nucleic acid binding protein-target protein based on the biochip of the present invention
[0039] Taking the transcription factor NFkB as the target protein, using pure NFkB, to detect the specificity of the method of the present invention based on biochip detection of whether there is an interaction between the nucleic acid binding protein-target protein, since this embodiment is to detect the specificity of the method, Among them, there is only one type of protein, NFkB, so there is no nucleic acid binding protein interacting with it. The specific detection method includes the following steps:
[0040] A. Preparation of DNA probes: the DNA chip-immobilized probes in each group were first dissolved in water and prepared in a 50% dimethyl sulfoxide (DMSO) solution with a final concentration of 10 μM. The nucleic acid-binding protein capture probes were all d...
Embodiment 2
[0047] Embodiment 2, detect the nucleic acid binding protein that interacts with protein STAT1 by the method of the present invention
[0048] Use the method of the present invention to detect whether there is an interaction between the nucleic acid binding protein-target protein (this embodiment is the transcription factor STAT1, GenBank number: NM_007315) based on the biochip to detect whether there is an interaction between the nuclear protein extracted after IFNγ stimulates the mouse 3T3L1 cell line and For nucleic acid binding proteins interacting with STAT1, the mixture of nucleic acid capture probes was added to the protein extract (the nuclear protein extracted after IFNγ stimulated the mouse 3T3L1 cell line in this example), and then passed through the antibody of the target protein (this example For example, the antibody of STAT1) separates the target protein and the nucleic acid binding protein interacting with it, and by detecting the nucleic acid sequence bound to ...
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