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38 results about "Fluorogenic Substrate" patented technology

Fluorogenic substrate. [′flu̇r·ə‚jen·ik ′səb‚strāt] (chemistry) A nonfluorescent material that is acted upon by an enzyme to produce a fluorescent compound.

Nano-enzyme-based ratiometric fluorescence sensing platform for determining total antioxidant capacity and application thereof

The invention relates to a nano-enzyme-based ratiometric fluorescence sensing platform for determining total antioxidant capacity and application thereof, and belongs to the technical field of preparation of fluorescence sensors. The ratio fluorescence sensing platform provided by the invention comprises MnSiO3 with oxidase-like activity, MnSiO3 with oxidase-like activity and MnSiO3 with oxidase-like activity, the invention relates to two fluorescent substrates with opposite responses. According to the ratio fluorescence sensing platform disclosed by the invention, the ratio fluorescence sensor is constructed by utilizing two fluorescent substrates, namely nano-enzyme MnSiO3 with oxidase-like activity and SC / OPD with opposite response, and is used for detecting TAC, so that the detection of three different antioxidants is realized. The ratio fluorescence sensing platform provided by the invention has the advantages of strong anti-interference capability and high sensitivity. The ratio fluorescence sensing platform has oxidase-like properties, does not introduce hydrogen peroxide, and has small interference on experiments.
Owner:OCEAN UNIV OF CHINA

Method for detecting glyphosate residues in coffee in dual modes of fluorescence and chemiluminescence

The invention discloses a method for detecting glyphosate residues in coffee in fluorescence and chemiluminescence dual modes, and belongs to the technical field of chemical analysis and detection.The method comprises the steps that succinic acid serves as a ligand, Ce < 4 + > serves as a central ion, a Suc-Ce complex is synthesized through a microwave method, then the Suc-Ce complex is further subjected to alkali treatment to obtain Suc-Ce-OH nano-enzyme, and the Suc-Ce-OH nano-enzyme is used for detecting glyphosate residues in coffee. The Suc-Ce-OH nano-enzyme has excellent phosphatase-like activity, surface coordinated water molecules can be effectively removed through alkali treatment, more Ce-O active sites are exposed, and the hydrolysis efficiency of the Suc-Ce-OH nano-enzyme on a fluorescent substrate 4-MUP and a chemiluminescent substrate AMPPD is improved by two times; based on the specific inhibition effect of glyphosate on phosphatase-like nano-enzyme activity, a glyphosate fluorescence and chemiluminescence dual-mode detection method is established, the detection limits of the glyphosate fluorescence and chemiluminescence dual-mode detection method are 0.05 mu g / mL and 0.01 mu g / mL respectively, and the glyphosate fluorescence and chemiluminescence dual-mode detection method is used for detecting glyphosate residues in a coffee sample and has the advantages of being high in specificity, easy to operate and sensitive.
Owner:YUNNAN AGRICULTURAL UNIVERSITY

Fluorogenic substrates for aminopeptidase detection in biofluids

The present application provides compositions and methods for determining a disease or condition in a subject. The method comprises contacting a body fluid with a molecule comprising a reporter thereof and the reported is cleaved by an agent in the body fluid. Diseases and conditions that can be determined by the methods are also described herein
Owner:SUNBIRD BIO INC

Method and application of a Cas12a sensing system based on sgRNA blocking strategy for single nucleotide polymorphism typing

The application discloses a method and application of a Cas12a sensing system based on an sgRNA blocking strategy for single nucleotide polymorphism typing. The method comprises the following steps: pre-annealing sgRNA and blocking chains to form a complex; obtaining a single-stranded DNA target by using an asymmetric recombinase polymerase amplification; constructing a detection system comprising a Cas12a protein, the complex, a target and a fluorescent substrate; activating the trans-cleavage activity of Cas12a through a strand displacement reaction to generate a fluorescent signal, so as to realize SNP typing. The method combines the sgRNA blocking strategy with the Cas12a system for the first time, does not need to depend on a PAM sequence, has the advantages of high universality, high specificity, simple operation, rapidness, low cost and the like, and is suitable for ApoE gene typing and other SNP related detection.
Owner:NANTONG UNIV

Fluorescent substrate 7-HC-P-E as well as preparation method and application thereof

The invention discloses a fluorescent substrate 7-HC-P-E, a preparation method thereof and application of the fluorescent substrate 7-HC-P-E in detection of the active concentration of alkaline phosphatase (ALP). The fluorescent substrate can be catalytically hydrolyzed by ALP under an alkaline condition to release 7-hydroxycoumarin (7HC) with a strong fluorescent signal, so that high-sensitivity and high-specificity detection on the enzyme activity is realized. The invention further provides a synthesis route and an optimization process of the fluorescent substrate, the operation is simple and convenient, the product purity is high, and the fluorescent substrate is suitable for multiple fields such as biological analysis, medical diagnosis and environmental monitoring.
Owner:HUAQIAO UNIVERSITY

Enzyme detection method and enzyme detection kit

InactiveJP2024179837AMicrobiological testing/measurementHexokinaseFluorogenic Substrate
To provide an enzyme detection method by which enzymes for synthesizing ATP or ADP can be detected, highly sensitive detection at one molecular level can be performed for an enzyme not having a fluorescent substrate among the enzymes, and highly sensitive measurement for a disease biomarker or the like can be performed; and an enzyme detection kit used in the same.SOLUTION: Provided are an enzyme detection method for detecting, from an enzyme sample, a target enzyme for synthesizing ADP or ATP, and an enzyme detection kit used in the same, the method comprising: a sample addition step for preparing a detection device including a microchamber array member in which a plurality of storage parts capable of being filled with a detection liquid are arranged, and filling the storage parts of the detection device with a detection liquid containing the enzyme sample and a hexokinase enzyme; and a detection step for detecting, from the detection liquid, a signal resulting from the synthesis of ADP or ATP.SELECTED DRAWING: None
Owner:THE UNIV OF TOKYO

A screening method for recombinant factor b activity and sensitivity for endotoxin detection

ActiveCN121431865BBiochemistryFluorogenic Substrate
The application discloses a screening method for activity and sensitivity of recombinant factor B for endotoxin detection. The method comprises the following steps: adding a series of endotoxin standard solution with different concentrations in an endotoxin-free 96-well plate, and adding endotoxin-free water as a control; adding an activity detection reagent, wherein the activity detection reagent comprises a buffer, a mixed solution of recombinant factor C and a to-be-detected recombinant factor B, and a fluorescent substrate; reading a zero-point fluorescence value, and then placing the 96-well plate in a 37 DEG C incubator for incubation; reading the fluorescence value again, calculating a fluorescence intensity change value Delta RFU, and determining the result. The method realizes independent and direct evaluation of the activity and sensitivity of the recombinant factor B through a 'two-factor' (only containing the recombinant factor C and the recombinant factor B) detection system, and completely gets rid of the dependence on downstream coagulase activity. The method can help screening of the recombinant factor B with high activity and sensitivity, and the recombinant factor B can be used in a recombinant limulus reagent for endotoxin detection.
Owner:BEIJING TRANSGEN BIOTECH CO LTD

Fluorogenic substrates and methods of use

The present disclosure relate to methods and fluorogenic substrates for analysis of analyte(s) in a sample. The methods include detecting presence of an analyte of interest on capture particles by contacting the capture particles present in nano-chambers with a fluorogenic substrate. The fluorogenic substrate may be of formula (I): (I) wherein: X is selected from a group consisting of H+, K+, Ca2+, tris, triethylammonium, or any other cation; Y is selected from a group consisting of diphosphate, triphosphate, oxymethyl phosphate, or phosphate.
Owner:ABBOTT LAB INC +8

Method for detecting activity of protease in yeast for making hard liquor

The invention discloses a method for detecting protease activity in yeast for making hard liquor. The detection method comprises the following steps: selecting carboxyl magnetic beads according to the characteristics of a Daqu sample, coupling protein to the magnetic beads to obtain a magnetic bead adsorption enzyme sample, marking casein by utilizing fluorescein isothiocyanate to prepare neutral and alkaline fluorescent substrates, and establishing a linear relation curve of enzyme activity and fluorescence intensity by utilizing neutral and alkaline protease standard substances. And finally, after a fluorescent substrate is hydrolyzed by protease to generate a soluble fluorescent peptide fragment, determining a fluorescence intensity value under a specific wavelength and substituting the fluorescence intensity value into the standard curve, so that the protease activity of the yeast for making hard liquor can be accurately determined. According to the detection method, the protein compound is adsorbed and separated through the magnetic beads, so that impurity interference in the yeast crude enzyme is effectively removed, the accuracy of enzyme activity determination is remarkably improved, and the protease activity of the yeast can be reliably determined.
Owner:SOUTH CHINA UNIV OF TECH

Microbiological detection guided water sample contamination test method

The present application relates to the technical field of water quality monitoring, and discloses a water sample pollution inspection method guided by microorganism detection, comprising: applying a pulsed electric field to a biochemical response system to reduce the mass transfer resistance of the cell membrane of an indicator microorganism by using a physical pore-forming effect; injecting a fluorescent substrate and synchronously collecting fluorescence emission intensity and backscattering light intensity; correcting the amplitude of the fluorescence emission intensity by using the backscattering light intensity to eliminate physical shielding interference and obtain a pre-steady-state fluorescence emission sequence; intercepting a characteristic value of the change rate of the pre-steady-state fluorescence emission sequence to calculate an intrinsic catalytic turnover index representing the specific catalytic activity of intracellular enzymes, and determining the pollution degree of the water sample to be tested according to the offset of the index relative to a preset reference value; and the present application can quickly identify pollutants with transmembrane mass transfer disorders and eliminate the influence of endogenous activity fluctuations of microorganisms on the detection result by means of the cooperation of electric pore-forming intervention and calculation logic.
Owner:江西省生态环境监测中心

Digital ELISA detection method to prevent fluorescence signal crosstalk

The present invention particularly relates to a digital ELISA detection method for preventing fluorescence signal crosstalk, the method comprising: preparing a sample containing a target molecule to be detected, capturing the target molecule to be detected using magnetic beads, and forming a complex of "magnetic beads-target molecule to be detected-enzyme" by affinity reaction; transferring the sample formed with the complex to a microwell array chip; transferring a fluorescent substrate to the microwell array chip; adding an isolation liquid to the microwell array chip; waiting for the enzyme to perform an enzymatic reaction with the fluorescent substrate to generate fluorescent molecules; determining the number of microwells containing magnetic beads in the microwell array and the number of microwells in the microwell array whose fluorescent signals are higher than a threshold value; based on the above two quantities, and based on the number of magnetic beads in the sample and the probability that the target molecule to be detected is captured by the magnetic beads and further combined with the enzyme, determining the number of target molecules to be detected in the sample. The present invention improves the accuracy of distinguishing negative signals and positive signals.
Owner:IOMICS BIOSCIENCES INC +1

Novel fluorescent substrates and uses thereof

The present disclosure relates to a novel fluorescent substrate, which is a compound or isomer of formula (I), and its use. A high-sensitivity detection reagent combination and detection method are also provided, which comprises the fluorescent substrate compound or isomer of formula (I) and an enzyme-labeled antibody. The fluorescent substrate and detection method can also be widely used for detecting cells with low expression of specific antigens. Compared with the traditional fluorescent secondary antibody detection method and the TSA-FITC combination method, the detection efficiency of the method of the present disclosure is higher, and the detection sensitivity and stability are higher.
Owner:ZHUHAI LIVZON CYNVENIO DIAGNOSTICS +1

Selective detection of fluorescent substrates and use in detecting s-comt activity

This invention discloses the selective detection of a fluorescent substrate and its application in detecting S-COMT activity, belonging to the field of analytical detection. The selective fluorescent substrate is 3-carboxy / methyldaphne (3A / M-DPN), which can be specifically catalyzed by S-COMT to generate an 8-O-methylated product, producing a strong fluorescence signal at 520 nm, and the fluorescence intensity shows a good linear relationship with S-COMT activity. The substrate of this invention has a high affinity for S-COMT (…). K m =1.41 μM) is much higher than that of membrane-bound MB-COMT ( K m =45.31 μM), catalytic efficiency ( V max / K m It is 280 times more potent than other substrates and possesses advantages such as high selectivity, high sensitivity, and ease of operation. This substrate can be used to distinguish the catalytic contribution of S-COMT from MB-COMT, quantitatively assess the S-COMT activity in various in vitro biological samples, and provide an important tool for S-COMT subtype functional studies, monitoring of S-COMT activity under pathological conditions, and screening of drugs targeting S-COMT.
Owner:SHANGHAI UNIV OF T C M

A method for dual-mode detection of glyphosate residues in coffee by fluorescence and chemiluminescence

The present invention discloses a method for detecting glyphosate residues in coffee by fluorescence and chemiluminescence dual-mode, belonging to the technical field of chemical analysis and detection. The method uses succinic acid as a ligand and Ce 4+ as the central ion, synthesizes the Suc-Ce complex by microwave method, and further obtains the Suc-Ce-OH nanozyme through alkali treatment. The Suc-Ce-OH nanozyme has excellent phosphatase-like activity. Alkali treatment can effectively remove surface coordinated water molecules and expose more Ce-O active sites, doubling its hydrolysis efficiency for the fluorescent substrate 4-MUP and the chemiluminescent substrate AMPPD. Based on the specific inhibitory effect of glyphosate on the activity of the phosphatase-like nanozyme, the present invention establishes a dual-mode detection method for glyphosate by fluorescence and chemiluminescence, with detection limits of 0.05 μg / mL and 0.01 μg / mL respectively. It is used to detect glyphosate residues in coffee samples, featuring strong specificity, simple operation, and high sensitivity.
Owner:YUNNAN AGRICULTURAL UNIVERSITY

DPP-IV (dipeptidyl peptidase-IV) inhibitory peptide screened from yak blood as well as screening method and equipment

The invention relates to the technical field of bioactive peptide screening, in particular to a DPP-IV inhibitory peptide screened from yak blood and a screening method and device.The method comprises the steps that yak blood is collected, blood cells are separated, a crude protein solution is obtained through crushing, centrifugation and fluorescent substrate preliminary screening, impurities are removed, marking is conducted, and a protein solution containing a fluorescent mark is prepared; specific polypeptide is fixed on an amino modified glass slide and incubated with a protein solution to prepare a DPP-IV inhibitory peptide solution containing a fluorescence label, signals are collected to extract a first peptide fragment set, sequence information of a second peptide fragment set is generated through sequence comparison and virtual enzymolysis, polypeptide is synthesized, and the inhibition effect is evaluated through incubation, fluorescence intensity detection and incubation in a culture medium. And an efficient and accurate method is provided for screening the DPP-IV inhibitory peptide.
Owner:NAQUGANG YAER ANIMAL HUSBANDRY TECHNOLOGY IND CO LTD

Method for screening activity and sensitivity of recombinant factor B for endotoxin detection

ActiveCN121431865ABiological testingCoagulogenBiochemistry
The invention discloses a method for screening activity and sensitivity of a recombinant factor B for endotoxin detection. The method comprises the following steps: adding a series of endotoxin standard substance solutions with different concentrations into an endotoxin-free 96-well plate, and taking endotoxin-free water as a contrast; adding an activity detection reagent, wherein the activity detection reagent comprises a buffer solution, a mixed solution of a recombinant C factor and a recombinant B factor to be detected, and a fluorescent substrate; reading a zero fluorescence value, and then placing the 96-well plate in an incubator at 37 DEG C for incubation; reading the fluorescence value again, and calculating a fluorescence intensity change value delta RFU; and judging a result. According to the method, independent and direct evaluation on the activity and sensitivity of the recombinant factor B is realized through a detection system of'two factors' (only containing the recombinant factor C and the recombinant factor B), and the dependence on the activity of the downstream clotting zymogen is thoroughly eliminated. The method can help to screen the recombinant factor B with high activity and sensitivity, and is used in a recombinant tachypleus amebocyte lysate for endotoxin detection.
Owner:BEIJING TRANSGEN BIOTECH CO LTD

Fluorescence-based AhR regulating agent screening method and application thereof

The invention provides a screening method of an aromatic hydrocarbon receptor (AhR) regulating agent based on fluorescence and application of the screening method. The detection principle is that the regulation effect of the small molecule on the AhR is quantitatively evaluated by detecting the activity of the AhR downstream cytochrome P450 1A1 (CYP1A1) through a biochemical method. Therefore, the invention firstly develops a specific fluorescent substrate suitable for functional imaging analysis of CYP1A1 in living cells. The structural general formula is as shown in formula (1): # imgabs0, the substrate can be specifically catalyzed by intracellular CYP1A1 under physiological conditions to perform dechloroethyl reaction and generate a single fluorescent product HDBI, the activity of CYP1A1 in living cells / living tissues can be quantitatively detected by detecting the generation amount of the fluorescent product HDBI in unit time, and then the agonistic activity of AhR is indirectly evaluated. A living cell-based CYP1A1 function detection method is constructed on the basis of the fluorescent substrate, and the fluorescent substrate can be used for high-throughput screening and evaluation of the regulation effect of a compound on an AhR-CYP1A1 axis so as to evaluate the regulation effect of the compound on a CYP1A1 upstream gene AhR.
Owner:SHANGHAI UNIV OF T C M

Immune cell population function evaluation and specificity screening method based on digital microfluidics

The invention discloses an immune cell population function evaluation and screening method based on digital microfluidics, which comprises the following steps: loading a cell suspension containing immune cells to an active matrix digital micro-fluidic chip, and controlling a chip electrode array to carry out on-chip segmentation and dilution on the cell suspension to form a plurality of cell droplets; segmenting a non-specific stimulant or a specific antigen injected into the chip, mixing the segmented non-specific stimulant or specific antigen with the cell droplets in proportion, and performing stimulation incubation; mixing the magnetic beads coated with the cytokine / immunoreaction secretion capture antibody and the enzyme-labeled cytokine / immunoreaction secretion detection antibody, and then mixing and incubating with the cell droplets; carrying out magnetic enrichment on the magnetic beads, and adding fluorescent substrate liquid drops for reaction; fluorescence intensity detection is carried out on the magnetic bead liquid drops, and positive and negative signals are identified, so that negative, strong-positive or weak-positive cell liquid drops after stimulation incubation are distinguished; positioning specific immune response positive cell liquid drops, screening and recycling, and / or continuously diluting and dispersing to complete specific immune cell screening.
Owner:SHENZHEN UNIV

Enzyme detection method and enzyme detection kit

PCT designated stage expiredWO2024257799A1Microbiological testing/measurementHexokinaseBiologic marker
Provided are: an enzyme detection method by which enzymes for synthesizing ATP or ADP can be detected, highly sensitive detection at one molecular level can be performed for an enzyme not having a fluorescent substrate among the enzymes, and highly sensitive measurement of a disease biomarker or the like can be performed; and an enzyme detection kit used in the same. Provided are an enzyme detection method for detecting, from an enzyme sample, a target enzyme for synthesizing ADP or ATP, and an enzyme detection kit used in the same, the method comprising: a sample addition step for preparing a detection device including a microchamber array member in which a plurality of storage parts capable of being filled with a detection liquid are arranged, and filling the storage parts of the detection device with a detection liquid containing the enzyme sample and a hexokinase enzyme; and a detection step for detecting, from the detection liquid, a signal obtained by synthesizing ADP or ATP.
Owner:THE UNIV OF TOKYO

Method and device for detecting urinary kidney injury molecule-1 protein by immunofluorescence

The application provides a urine kidney injury molecule-1 protein immunofluorescence detection reagent detection method and device, is applied to the biomedical detection technical field, through pretreatment to the sample, obtains the dilution urine sample, and then adjusts the pH range of the dilution urine sample, and then utilizes the specific recognition of the antibody to the antigen to form a complex, uses the capture antibody fixed in the detection area in advance to capture the immune complex, makes the immune complex enrich in the detection area, improves the detection signal. Drop the fluorescence substrate to the detection area, finally use the portable fluorescence detector to detect the fluorescence intensity of the detection area, obtain the concentration of the urine kidney injury molecule-1 protein in the urine sample, realize quantitative detection. Therefore, the application realizes the rapid, simple and quantitative detection of the urine kidney injury molecule-1 protein in the urine sample.
Owner:JIANGSU MEIDE PHARM TECH CO LTD

A green preparation method for magnetic particle enhanced biomolecule detection without surface modification

A method for green preparation of magnetic particle enhanced biomolecule detection without surface modification belongs to the field of biomolecule detection methods, and includes the following steps: adding metal iron salt and metal copper salt into a chloroplast solution; adding NaOH to adjust the pH value and stirring for reaction; centrifuging to collect the precipitate; washing; calcining; adding copper ferrite particles and a capture antibody corresponding to the biomolecule to be detected into a centrifuge tube; washing away the uncaptured capture antibody; adding a blocking solution; washing; adding the biomolecule to be detected; washing away the unreacted biomolecule to be detected; adding a detection antibody; washing away the unreacted detection antibody; adding streptavidin labeled with alkaline phosphatase; washing away the uncrosslinked streptavidin; adding a chemiluminescent substrate; adding a termination solution and detecting the absorbance. The present invention solves the problem that the magnetic beads prepared by the existing method cannot be directly applied to ELISA due to biological toxicity and lack of active groups on the surface and need further biochemical surface modification.
Owner:TAIYUAN UNIVERSITY OF TECHNOLOGY

CRISPR-Cas12a (clustered regularly interspaced short palindromic repeats-associated 12a)-based nucleic acid detection kit as well as preparation and application methods thereof

The invention relates to the technical field of nucleic acid molecule detection, and discloses a multiple nucleic acid detection kit based on CRISPR-Cas12a, the multiple nucleic acid detection kit comprises a plurality of fluorescence coding microspheres, and each fluorescence coding microsphere comprises a lipid membrane coated solid phase carrier microsphere, a lipid membrane coated solid phase carrier microsphere, a lipid membrane coated solid phase carrier microsphere and a lipid membrane coated solid phase carrier microsphere, the Cas12a / crRNA compound is embedded into the lipid membrane through hydrophobic interaction, and the 3'end of the crRNA is covalently connected with a cholesterol group; the fluorescent substrate probe is embedded into the lipid membrane through hydrophobic interaction and is single-stranded DNA, the 3'end of the fluorescent substrate probe is covalently connected with a cholesterol group, and the 5 'end of the fluorescent substrate probe is covalently connected with a fluorescent group. Different crRNA / Cas12a compounds and fluorescent substrate probes are jointly anchored on the surfaces of the microspheres with different optical codes, each microsphere corresponds to one specific target, multiple nucleic acid targets can be simultaneously detected in the same reaction system, and the detection flux and efficiency are remarkably improved.
Owner:HEFEI UNIV OF TECH

Method for analyzing intracellular glycosylated RNA (Ribonucleic Acid) by modifying retardation nucleic acid hybridization technology

The invention discloses an intracellular glycosylated RNA (Ribonucleic Acid) analysis method for modifying a retardation nucleic acid hybridization technology. Analysis detection and in-situ imaging of intracellular glycosylated RNA (glycoRNAs) are realized on the basis of structural transformation of a target nucleic acid mediated fluorescent substrate nucleic acid chain. Specifically, a substrate chain with toehold is designed according to a sequence of microRNA-21 and a glycosylation modification site of the microRNA-21, and the sequence of the substrate chain contains a specific sequence of a Pepper aptamer capable of being combined with a fluorescent small molecule dye HBC. And the sequence design of the substrate meets the condition that when the substrate is hybridized with glyco-microRNA-21 to a glycosylation modification site, the structure of the generated hybridized compound can just generate an 'ON' fluorescence signal. The hybridization of the substrate sequence and the microRNA-21 is full hybridization, and the formed hybridization product cannot generate a fluorescence signal. According to the invention, the detection of low-abundance glycoRNAs under the interference of a large number of non-glycosylated RNAs is realized, and the intracellular in-situ imaging of the glycoRNAs is realized.
Owner:SOUTHWEST UNIV

A compound, composition and use

PendingCN122647465AChemical compoundFluorophore
The application discloses a compound, a composition and application, and claims the compound shown in formula (I) or a salt thereof. The compound provided by the application is an enzyme responsive fluorescent substrate specially designed for sulfotransferase SULT1A1, and precise recognition and specific catalytic response to SULT1A1 are realized from the molecular structure level. The compound has a low fluorescent background itself, the sulfonated fluorophore in the catalytic reaction product has the characteristics of near-infrared emission and large Stokes shift, can effectively avoid the background interference of excitation light on fluorescent detection, and is excellent in specificity, stability and linearity. Therefore, the compound or the salt thereof and the composition containing the compound or the salt thereof have the prospect of being developed into a reagent, a kit or a microplate for in vitro quantitative or qualitative detection of SULT1A1 activity, in situ imaging and positioning of SULT1A1 in cells or in vivo, and high-throughput screening of SULT1A1 inhibitors or activators.
Owner:CHINA PHARM UNIV

A fluorescence-based AhR regulator screening method and its application

The present invention provides a fluorescence-based screening method for aryl hydrocarbon receptor (AhR) modulators and its application. The detection principle is to quantitatively evaluate the regulatory effect of small molecules on AhR by biochemically detecting the activity of cytochrome P450 1A1 (CYP1A1) downstream of AhR. To this end, the present invention first developed a class of specific fluorescent substrates suitable for CYP1A1 functional imaging analysis in living cells. Its general structural formula is shown in formula (1): This type of substrate can be specifically catalyzed by intracellular CYP1A1 under physiological conditions to undergo a dechloroethylation reaction and produce a single fluorescent product HDBI. The activity of CYP1A1 in living cells / living tissues can be quantitatively detected by detecting the amount of fluorescent product HDBI generated per unit time, thereby indirectly evaluating the agonist activity of AhR. Based on this type of fluorescent substrate, a living cell-based CYP1A1 functional detection method is constructed, which can be used for high-throughput screening and evaluation of the regulatory effect of compounds on the AhR-CYP1A1 axis, and further evaluate the regulatory effect of compounds on the CYP1A1 upstream gene AhR.
Owner:SHANGHAI UNIV OF T C M

Kit for integrally detecting senescent cells and apoptotic cells as well as detection method and application of kit

The invention provides a kit for integrally detecting aging cells and apoptotic cells as well as a detection method and application of the kit. The kit comprises an aging inducer, an apoptosis inducer, an integrated buffer solution, a beta-galactosidase specific fluorescent substrate and an apoptotic cell fluorescent staining solution. The kit disclosed by the invention contains an integrated buffer solution capable of fully meeting the detection requirements of senescence and apoptosis cell populations and transition cell populations, so that the experimental steps can be simplified to the greatest extent, and the experimental cost is saved. Results provided by the kit are visual, data are accurate, the detection accuracy and sensitivity are remarkably improved, and a powerful tool is provided for integrally analyzing and detecting dynamic evolution of the aging process and the apoptosis process through a streaming technology.
Owner:TIANJIN MEDICAL UNIV

Simoa triple detection reagents, kits and applications for biomarkers IL-8, hCGβ and RANTES

The present invention relates to Simoa triple detection reagents, kits and applications of biomarkers IL-8, hCGβ and RANTES, and to the field of immunoassay technology. The Simoa triple detection reagent includes capture antibody-coated magnetic beads, biotinylated detection antibodies, streptavidin-β-galactosidase solution and fluorescent substrate solution; the magnetic beads coated with the capture antibody of IL-8, the magnetic beads coated with the capture antibody of hCGβ and the magnetic beads coated with the capture antibody of RANTES are magnetic beads with different fluorescence encodings. The Simoa triple detection reagent or kit of the present invention realizes high-throughput detection of multiple protein markers that can evaluate the potential of embryo implantation, realizes simultaneous detection of multiple protein markers in one detection, and provides the possibility for analyzing and studying the relationship between the target protein in the conditioned culture medium of a single blastocyst and the embryo quality and the pregnancy outcome of the transplantation.
Owner:SHENZHEN ZHONGSHAN OBSTETRICS & GYNECOLOGY HOSPITAL

Anti-dextran interference endotoxin detection method and anti-dextran interference endotoxin detection system

The invention discloses an anti-dextran interference endotoxin detection method and system, and belongs to the technical field of detection and analysis of bioactive substances, and the method comprises the following steps: obtaining a dynamic fluorescence signal of a reaction mixture after a sample to be detected is mixed with a specific reaction system containing a recombinant C factor enzyme solution and a fluorescent substrate, and generating a dynamic baseline model; carrying out feedback incubation control on the reaction mixture according to the dynamic baseline model and the functional model for indicating the standard dynamic behavior, generating actual dynamic data, and carrying out multi-dimensional signal fusion calculation in combination with the dynamic baseline model to generate a fusion fluorescence signal difference value; and performing self-verification concentration judgment according to the fused fluorescence signal difference value and the curve family for indicating the relationship between the endotoxin and the fluorescence response, and generating the endotoxin concentration of the sample. By adopting a dynamic baseline model, feedback incubation control, multi-dimensional signal fusion calculation and self-verification concentration judgment, non-specific interference such as glucan can be eliminated, and the accuracy and reliability of an endotoxin quantitative result are ensured.
Owner:SUZHOU RUITEBAI BIOTECHNOLOGY CO LTD

Phosphor wheel and light emitting device

A phosphor wheel (1B) that is transmissive, and includes: a fluorescent substrate (10B) that consists essentially of a fluorescent material; a metal member (20B) bonded to a principal surface that the fluorescent substrate (10B) includes, the metal member (20B) including a plurality of first radiator fins (202) that protrude in a direction away from the principal surface; and a rotator (30B) that rotates the fluorescent substrate (10B) and the metal member (20B) about an axis extending in a thickness direction of the fluorescent substrate (10B). In the phosphor wheel (1B), in a plan view of the fluorescent substrate (10B), the fluorescent substrate (10B) includes an annular ring-shaped region (A11) that does not overlap the metal member (20B), and in a plan view of the metal member (20B), an opening (206) is provided between the plurality of first radiator fins (202) of the metal member (20B).
Owner:PANASONIC INTELLECTUAL PROPERTY MANAGEMENT CO LTD