Method for carrying out enzyme-linked immunoadsorption detection on integral zebra fish and application thereof
An enzyme-linked immunosorbent assay and zebrafish technology, applied in biological testing, material inspection products, etc., can solve the problems that cannot reflect the real situation of compounds, cannot perform high-throughput screening, and have many false positive results, so as to facilitate drug delivery, The administration method is safe and the effect is reproducible
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Embodiment 1
[0172] Example 1 Determination of the optimal developmental stage of zebrafish during compound treatment
[0173] Embryo collection
[0174] In the early morning, 5 pairs of zebrafish were taken, freely combined and divided into 5 groups to mate and hatch embryos, and 1200 zebrafish were collected, the sediment was sucked out, put into the incubation solution, and incubated in a 28°C incubator. Take out the white embryos (dead) in time to prevent the water quality from deteriorating. During this period, the zebrafish obtains nutrients through the yolk, so no other nutrients are needed.
[0175] drug treatment
[0176] The zebrafish were divided into 7 experimental groups, which were 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, and 7 days after fertilization. Each experimental group was further divided into three groups, blank group, 0.1% dimethyl sulfoxide (DMSO) negative control group and experimental treatment group (50 μM staurosporine, 0.1% dimethyl sulfoxide (DMSO) D...
Embodiment 2
[0184] Example 2 Determination of optimal time length for compound treatment
[0185] Embryo collection
[0186] In the early morning, 5 pairs of zebrafish were taken, freely combined and divided into 5 groups to mate and hatch embryos, and 1200 zebrafish were collected, the sediment was sucked out, put into the incubation solution, and incubated in a 28°C incubator. Take out the white embryos (dead) in time to prevent the water quality from deteriorating. During this period, the zebrafish obtains nutrients through the yolk, so no other nutrients are needed.
[0187] drug treatment
[0188] The zebrafish were divided into five experimental groups, each group including blank group, treatment group (50 μM staurosporine (staurosporine), dissolved in 0.1% dimethyl sulfoxide (DMSO)) and negative control group (0.1% dimethyl sulfoxide (DMSO) sulfoxide (DMSO)). The experimental groups were treated 2 days after zebrafish fertilization, and the treatment time was 6 hours, 12 hours, ...
Embodiment 3
[0196] Example 3 Detection of drug genotoxicity
[0197] Embryo collection
[0198] In the early morning, 10 pairs of zebrafish were taken, freely combined and divided into 10 groups to mate and hatch embryos, 1200 zebrafish were collected, the sediment was sucked out, put into the incubation solution, and incubated in a 28°C incubator. Take out the white embryos (dead) in time to prevent the water quality from deteriorating. During this period, the zebrafish obtains nutrients through the yolk, so no other nutrients are needed.
[0199] drug treatment
[0200] The zebrafish were divided into eight experimental groups, respectively 0.1, 1, 10, 100, 1000 μM genotoxic drug cisplatin (cisplatin), and then dissolved in the incubation solution according to the above concentration; the blank group was no treatment, 0.1% Dimethyl sulfoxide (DMSO) negative control group, 50μM staurosporine (staurosporine) positive control group that can cause genotoxicity. Processing starts at 2dpf ...
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