402 results about "Blot" patented technology

Relating to sensors and molecular imprinting technologies, the invention discloses an electropolymerization molecular imprinting technology-based double-parameter composite micro-sensor and a preparation thereof. According to the invention, three electrochemical microelectrode systems are integrated on a same chip, and each electrochemical microelectrode system has its independent micro-electrochemical reaction pool. Through encapsulation by a sealant, the integrated chip can form an open composite measurement pool containing three electrochemical microelectrode systems. In the micro-reaction pool of each electrochemical microelectrode system, by injecting a solution from the outside, a molecular imprinting procedure containing in situ polymerization and ultrasonic elution of template molecules can be implemented separately, thus obtaining a molecularly imprinted sensor. The left and right microelectrode systems of the composite micro-sensor respectively recognize two corresponding molecules due to different molecularly imprinted sensitive membranes, and the electrochemical microelectrode system positioned in the middle is used as a differential detection reference so as to deduct background signals and environmental effects of a test system. The composite micro-sensor provided in the invention can recognize two molecules simultaneously.

The invention relates to a molecular imprinting chemiluminescence sensor for detecting trace amount pesticide residues, belonging to the technical field of the detection of the pesticide residues. Theinvention also relates to a method for detecting the trace amount pesticide residues contained in agricultural product samples by using the molecular imprinting chemiluminescence sensor, which comprises the following steps: (1) selecting a functional monomer; (2) uniformly mixing a template molecule, the functional monomer, a cross linker, a pore-forming agent, an initiating agent and a silicon source to prepare an MIPs collosol according to a mole ratio of 0.1-1 to 1-10 to 5-25 to 20-60 to 0.05-0.15 to 20-50; (3) preparing a quantum dot material solution; and (4) modifying the MIPs collosoland quantum dot materials to the surface of a millipore plate to manufacture the chemiluminescence sensor by a layer-layer self-assembly surface modification technique. The invention has the advantages of high specificity and sensitivity, fast detection, low cost, good repeatability and convenient field detection on the detection of the pesticide residues.

Systems and methods are provided for integrating the electrophoresis and blotting of samples. A capillary electrophoresis and blotting system allows concomitant electrophoretic separation and blotting to provide a rapid and simplified process. A microfluidic electrophoresis and blotting system provides electrophoretic separation in a microfluidic channel followed by electroblotting from the microfluidic channel to also provide a rapid and simplified process. These systems and methods can be used to assay smaller amounts of sample in less time than conventional processes, including conventional Western blotting techniques.

The invention provides a magnetic molecularly imprinted nano-particle as well as a preparation method and an application thereof. The nano-particle can perform specific recognition, trapping, separation and activity inhibition on target protein in a solution in vitro, more importantly, the nano-particle can enter a living cell rapidly under the action of a magnetic field and perform in-situ combination and activity inhibition on the target protein in the living cell, distribution of the nano-particles in the cell can be traced through a fluorescence microscope after fluorescence labeling, and quantitation can be performed through detection of fluorescence intensity.

The invention discloses an E.tenella protein disulfide linkage isomerase gene EtPDI (Clone ID is BW1-E06,and the Genbank accession number of is EF552214). The gene is connected with a procaryon expression vector pGEX-4T-2; a procaryon expression recombination plasmid pGEX-4T-EtPDI is constructed and is expressed in a colibacillus system; and most of the expressed recombining protein exists in a soluble form. The recombining protein 4T-EtPDI is purified to carry out SPS-PAGE and is transferred to a PVDF film; and antiserum of E.tenella oocyst oral immunized chicken is used as first resistance and goat anti-chicken IgG is used as second resistance to carry out Western-blot analysis, thereby indicating that the gene has certain antigen. The gene is used for preparing an anti-chicken coccidiosis drug and an anti-chicken coccidiosis vaccine.

The invention provides a high-specificity high-sensitivity coordinated compound anti-human-TK1 antibody prepared from an antigenic determinant composed of human cervical cancer cell TK1 monomer N-terminal 23 peptide, C-terminal 20 peptide and C-terminal 28 peptide, and application thereof in tumor diagnosis. The antigenic determinant contains the following amino acid sequences: N-terminal 23 peptide (3-25): CINLPTVLPGSPSKTRGQIQVIL, C-terminal 20 peptide (206-225) CPVPGKPGEAVAARKLFAPQ, and C-terminal 28 peptide (198-225) AGPDNKENCPVPGKPGEAVAARKLFAPQ. The invention also relates to a method for preparing the antibody prepared from the antigen. The antibody kit provided by the invention has the characteristics of high sensitivity, high specificity, low cost and the like. The treatment effect on the tumor patient is evaluated by an enhanced chemiluminescent point blotting detection method, immunohistochemical detection and the detection kit.

The invention provides a high-specificity and high-sensitivity coordinated antibody against human thymidine kinase 1 (TK1) prepared from an antigenic determinant consisting of 23 peptides at an N terminal, 20 peptides at a C terminal and 28 peptides at the C terminal of the TK1 monomer from a human hela cell and the use of the detection and diagnosis system of the antibody in tumor diagnosis. The antigenic determinant comprises the following amino acid sequences: the sequence of the 23 peptides (3-25) at the N terminal:CINLPTVLPGSPSKTRGQIQVIL; the sequence of the 20 peptides(206-225) at the C terminal: CPVPGKPGEAVAARKLFAPQ; and the sequence of the 28 peptides (198-225) at the C terminal: AGPDNKENCPVPGKPGEAVAARKLFAPQ. The invention also provides a method for preparing the antibody by using the antigen. An antibody kit provided by the invention has the characteristics of high sensitivity, high specificity, low cost and the like; and the early tumor can be detected and pre-warned in mass physical examination screening by enhanced chemiluminescence dot blot assay, immuno-histochemistry and the detection kit.

According to the invention, the culture time of paddy rice is shortened by adjusting the growing conditions of paddy rice seedlings; a paddy rice green protoplast is prepared by an enzyme method; the preparation method is optimized, so vacuum pumping and the usage of a highly toxic reagent are avoided; and thus the preparation method of the paddy rice green protoplast is simpler and safer. The protoplast prepared by the method of the invention has a high cell survival rate, and a high transformation rate, is applicable to various analysis such as subcellular localization of target gene codingprotein, protein interaction, western blotting and the like, and is especially applicable to instantaneous expression research of light/chloroplast related gene.

The invention discloses a preparation method of a blotting membrane for Hg<2+> detection. The preparation method comprises the following steps: designing and synthesizing Rhodamine derivative with special developing function to Hg<2+>; and opolymerizing on a support polyvinylidene fluoride membrane by using target metal ion as a template, ethylene dimethacrylate as a crosslinking agent, azodiisobutyronitrile as a radical initiator, and the synthesized Rhodamine derivative as a function monomer to obtain the blotting membrane for mercury ion detection. The characteristic that the ionic blotting technology has high selectivity to the template ion is adopted by the invention to combine with the simplicity of a colorimetric/fluorescence chemical sensor, a colorimetric/fluorescence chemical sensing blotting membrane with high selectivity to Hg<2+> is prepared, and a fast and high-speed new scene detection method with high sensitivity and high selectivity is hopefully provided for heavy metal ions in environment and food.

A method of determining time of death is disclosed. Cardiac troponin I (cTnI) is the most specific marker for the determination of myocardial infarction (MI). Cardiac troponin I has a distinctive temporal degradation profile after death, which is key to its use as a time of death marker in forensic medicine. The method consists of sampling cardiac tissue, extracting a cardiac protein from a homogenate via methods which may include the magnetic microparticles or magnetic microparticles linked to anti-cTnI antibodies, eluting the proteins, separating proteins by electrophoresis, transferring by western blot the different bands of proteins to paper. A comparison/analysis of the concentration and kinetics of degradation of the protein at a given temperature(s) against known standards after time of death provides an accurate measurement of the time of death. The present invention is more accurate and reliable than the rudimentary time of death techniques currently used by medical examiners.

The invention relates to a test paper strip for rapidly detecting traces of chlorothalonil and a preparation method thereof. The test paper strip is characterized in that a supporting layer is used as the bottom layer, an adsorption layer is used as the intermediate layer, a protective layer is fixed on the adsorption layer, the adsorption layer comprises an adsorption fibrous layer, a gold-labeled antibody fibrous layer, a cellulose film and an absorbent material layer at the handle end in order from a test terminal, wherein the cellulose film is provided with detection blots printed by using a chlorothalonil coupling carrier protein solution and control blots printed by using a goat anti-mouse IgG or rabbit anti-mouse IgG (or goat anti-rabbit IgG) antibody solution; the gold-labeled antibody is a colloidal gold labeled chlorothalonil monoclonal antibody or polyclonal antibody; and the chlorothalonil coupling carrier protein is bovine serum albumin, chicken ovalbumin or haemocyanin. The test paper strip disclosed herein has the advantages of strong specificity, high sensitivity, simple, and accurate detection, low cost, wide application scope, and easiness in popularization and application.

The invention discloses a chip for diagnosing the proteins. The antigens corresponding to multiple relevant indexes are printed on the different positions of the membrane of cellulose nitrate. The multi-tape immunoblotting method is utilzied through the coloration of gold mark or the precipitation enzyme without coloration to detect the antibody in the specimen of the sufferer aiming at the antibodies all antigens. The invention raises the detecting sensitivity and the specificity of the immunizing method so as to avoid the false negative or the false positive, providing the results for detecting multiple antigens in one one-stroke detection. Since the chip possesses more information than the information in the testing sheet so as to reduce the collecting quantity and shorten the testing period.

The invention provides a high-specificity and high-sensitivity coordination combination anti-human TK1 antibody prepared from an antigenic determinant consisting of 23 peptide at an N end, 20 peptide at a C end and 28 peptide at the C end of human cervical cancer cells, and application thereof to tumor diagnosis. The antigenic determinant comprises the following amino acid sequences: 1) the 23 peptide (3-25) at the N end: CINLPTVLPGSPSKTRGQIQVIL: 2) the 20 peptide (206-225) at the C end: CPVPGKPGEAVAARKLFAPQ; and 3) the 28 peptide at the C end: AGPDNKENCPVPGKPGEAVAARKLFAPQ. The invention simultaneously provides a method applying the antigen for preparing the antibody. An antibody kit provided by the invention has the characteristics of high sensitivity, high specificity, low cost and the like, tumor rehabilitation populations are periodically monitored by an enhanced chemiluminescence dot blot detection method, immunohistochemistry detection and the detection kit, and the recurrence and transfer risk and the prognosis of the tumor rehabilitation people are evaluated.

The invention relates to an immunochromatography test strip marked by phosphorescent silica nano particles and used for quantitatively detecting cimaterol and a preparation method for the immunochromatography test strip. The test strip comprises a supporting layer, an adsorption layer and a protecting layer, wherein the adsorption layer sequentially comprises an adsorption fiber layer, a phosphorescent antibody fiber layer, a cellulose membrane layer and a water absorbing material layer at a handle end from a testing end; invisible detecting blotting printed by a carrier protein solution of coupling cimaterol and invisible comparison blotting printed by an anti-goat or anti-rabbit mouse IgG antibody solution are arranged on the cellulose membrane layer; the phosphorescent antibody fiber layer is made from glass cellucotton adsorbing phosphorescent antibody; and the phosphorescent antibody is cimaterol antibody marked by the phosphorescent silica nano particles. The test strip is high in specificity and sensitivity, is simple, convenient, visualized and accurate, can be used for quantitative and qualitative detection, can detect pg-scale trace residues to the minimum degree, and is wide in application range, low in cost and easy to popularize and apply.

A composite magnetic gel microsphere for biological macromolecualr template blot, based on the template blot (molecular blot) technique, is prepared from different biological macromolaculate as template molecule and different metals (or their oxides) as magentic component through reversed-phase suspension polymerization. It can be used for separating biological macromoleculae with high precision and simple process.

The invention provides a preparation method of mythyl p-hydroxybenzoate molecular imprinting composite membrane and application thereof. A molecular imprinting composite preparation method and an application of mythyl p hydroxybenzoate in separation of methyl salicylate belong to the fields of material preparation technology and separating technology. polyvinylidene fluoride micro filter membrane is used as a substrate, and mythyl p-hydroxybenzoate (MP) which is impurity in methyl salicylate is used as a template molecule, and methacrylic acid (MAA) is used as a functional monomer, and N, N'-methylene bisacrylamide (MPAA) is used as a cross-linking agent, and azodiisobutyronitrile (AIBN) is used as an initiator; preparation of mythyl p-hydroxybenzoate molecular blotting membrane (MIMMP), static adsorption experiment and selective penetration experiment are used for researching selective identification property of the preparative molecular imprinting membrane. The result shows that, according to the invention, the obtained mythyl p-hydroxybenzoate molecular imprinting composite membrane has super mythyl p-hydroxybenzoate molecule identification performance.

The invention discloses an expression process of a coat protein gene of prunus necrotic ring spot virus (PNRSV), an antiserum and a kit. The expression method comprises the following steps of: extracting a ribose nucleic acid (RNA) of the PNRSV in an affected material; adopting a transcription-polymerase chain reaction (RT-PCR) amplification process to clone the coat protein gene of the PNRSV; connecting the obtained coat protein gene to a pGEM-T vector, obtaining a pGEM-T-PNRSV recombinant vector, and using sequence analysis to determine the sequence accuracy of the coat protein gene of the PNRSV; building an eukaryotic expression vector of the coat protein gene of the PNRSV; transforming a pichia pastoris GS115 strain, and inducing a pichia pastoris expression coat protein; and using Western-blot to detect the size accuracy of an expression proteine. The invention aims to provide the process for using the pichia pastoris to express the PNRSV coat protein gene, the antiserum, and a PNRSV ELISA detection kit.

The invention discloses a gold-labeled test strip for rapid detection of chromium ions as well as a preparation method and an application thereof. The gold-labeled test strip is characterized in that a bottom layer is a support layer, a middle layer is an absorption layer, a protection film is fixed on the absorption layer, the absorption layer is sequentially provided with a sample pad, a gold-labeled antibody binding pad, and a cellulose membrane layer from a test end as well as a water absorption pad at a handle end, detection prints printed by a carrier protein solution coupled with the chromium ions are arranged on the cellulose membrane layer, and contrast prints printed by rabbit-anti-mouse or goat-anti-mouse IgG (Intravenous Gamma Globulin) antibody solution are arranged on the cellulose membrane layer; colloidal gold-labeled chromium ion monoclonal antibodies are coated in the gold-labeled antibody binding pad. The gold-labeled test strip can be used for rapidly detecting pollution residues of the chromium ions in soil, water and food, has the advantages of specificity, sensitivity, rapidness, simplicity, convenience, visual and intuitional result and the like, not only can be used for screening large-batch samples, but also can be used for rapidly detecting small-batch samples and is wide in applicable range.

The present invention relates to a classical swine fever antibody PPA-ELISA detection kit and a preparation method thereof. The kit preparation comprises: adopting RT-PCR to amplify a 550 bp gene fragment of E2 gene, wherein the fragment contains 4 neutralizing antigen regions of A, B, C and D; cloning the amplified 550 bp gene fragment into a pMD18-T vector; carrying out identification, wherein the identification result is correct; further directionally cloning the fragment into a PGEX-6P-1 prokaryotic expression vector; transforming BL21 expression bacterial; carrying out IPTG induced expression to obtain recombinant protein expressed in a form of inclusion body; adopting an affinity chromatography method to separate and purify the recombinant protein; adopting immunoblotting to detect antigenicity and specificity of the purified protein; and adopting the recombinant protein as coating antigen, adopting horseradish peroxidase-labeled staphylococcal protein A (SPA) as second antibody, optimizing reaction conditions of indirect ELISA, establishing an indirect ELISA detection method for detection of classical swine fever virus antibody, and assembling the classical swine fever antibody PPA-ELISA detection kit.

The invention discloses a ratiometric fluorescent molecularly imprinted polymer and a preparation method and application thereof. Sol-gel polymerization imprinting is performed on the surfaces of thesilicon dioxide nanoparticles through a one-step method; c-type natriuretic peptide is used as a template molecule, 3-aminopropyltriethoxysilane is used as a functional monomer, tetraethyl orthosilicate is used as a cross-linking agent, two APTES modified fluorescent nano-material carbon dots and nitrobenzoxazole are grafted at the same time, the template molecule is eluted, and the ratiometric fluorescent molecularly imprinted polymer nanoparticles are obtained. The preparation of the ratiometric fluorescent molecularly imprinted polymer is optimized through a one-step method, the consumed time is short, and the process is simple. The ratiometric fluorescent molecularly imprinted polymer synthesized by the method has high affinity and selectivity to imprinted molecules, is low in preparation cost, stable in process and low in energy consumption, can be used for efficiently separating and detecting C-type natriuretic peptide in a biological sample, and is expected to solve the problemthat a biomarker is difficult to separate and detect.