Integrated micro-fluidic chip used for fluorescence detection of enzyme catalysis product and application thereof
A microfluidic chip and fluorescence detection technology, applied in the direction of fluorescence/phosphorescence, laboratory containers, chemical instruments and methods, etc., can solve the problems of affecting enzyme-catalyzed reactions, changing reactions, and difficulty in obtaining fluorescent substrates, etc., to achieve Improve sensitivity, reduce analysis errors, and avoid mutual interference effects
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Example Embodiment
[0031] Example 1
[0032] The needles of the dual-channel micro-injection pump are respectively connected with the chip input ports C1 and C2 connecting pipelines, the input ports C3 and C4 are opened, and the liquid flows out of the chip through the input ports C3, C4 and the waste liquid port C5. Rinse the channel with 0.1M NaOH for 30min, rinse with distilled water and then rinse with 0.1M HCl for 30min, and finally rinse the chip channel with 0.01PBS buffer.
[0033] After completing the above-mentioned channel flushing, the input ports C3 and C4 are closed, and the liquid flows out of the chip from the waste liquid port C5. Through the input ports C1 and C2, BSA (1mg / mL) was input into the lower microchannel network of the chip at a constant flow rate of 10μL / min, and left to stand for 3.5 hours. Then, the mixture solution of protein A (20μg / mL) and 1% glutaraldehyde, rabbit immunoglobulin IgG (5μg / mL dissolved in 0.01M PBS) solution was sequentially input, and incubated for ...
Example Embodiment
[0035] Example 2
[0036] Fix the urease in the channel of the microfluidic chip according to the operation steps described in Example 1. The chip is placed in a constant temperature bath at 37°C, and the needles of the two dual-channel microinjection pumps are connected to the input ports C1 and C2 on the chip respectively; C3 , C4 connecting pipeline is connected. From the input ports C1 and C2, two 0.2M phosphate buffer solutions with different concentrations of urea (0mM, 0.5mM) were input into the lower microchannel network at a constant flow rate of 10μL / min. The concentration gradient generating unit generates 6 rows of urea solutions with different concentrations to contact the immobilized urease on the channel to cause an enzyme-catalyzed reaction. After incubating for 3 minutes, urea is hydrolyzed by enzymes to produce ammonia, which is carried to the upper microchannel network through 6 communicating holes. At the same time, the OPA reagent (that is, the fluorescence...
Example Embodiment
[0037] Example 3
[0038] Fix the urease in the channel of the microfluidic chip according to the operation steps described in Example 1. The chip is placed in a constant temperature bath at 37°C, and the needles of the two dual-channel microinjection pumps are connected to the input ports C1 and C2 on the chip respectively; C3 , C4 connecting pipeline is connected. From the input ports C1 and C2, two different concentrations of acetyl hydroxamic acid (0mM, 0.1mM) and 0.2M phosphate buffer solution of the same concentration of urea (0.1mM) were injected into the lower microchannel at a constant flow rate of 10μL / min The internet. The concentration gradient generating unit generates 6 columns of solutions with different concentrations of acetyl hydroxamic acid and the same concentration of urea, which contact the immobilized urease on the channel to produce an enzyme catalytic reaction, and at the same time, acetyl hydroxamic acid inhibits urease. After incubating for 3 minutes,...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap