425 results about "Blot" patented technology

The invention describes methods for labeling or detecting of immobilized poly(amino acids), including peptides, polypeptides and proteins, on membranes and other solid supports, using fluorescent dipyrrometheneboron difluoride dyes. Such immobilized poly(amino acids) are labeled or detected on blots or on arrays of poly(amino acids), or are attached to immobilized aptamers.

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.

A rapid, efficient and convenient method to detect one or more biological entities on a blotting membrane is provided. The detection can relate to the position, nature or amount of the biological substance on one or more membranes. The invention method involves a pressure assisted regiment (such as vacuum or positive gas pressure) for the supply and removal of reagents and permits washing of the contaminants from substances embedded in the membrane to be detected using very low volumes of liquid. This method enables completion of the blocking, washing and antibody binding steps in about 30 minutes without comprising blot quality. In another aspect, the invention is directed to an apparatus useful in conducting the method of the invention. The device is comprised of several layers including a porous support layer below the blotting membrane(s), a flow distributor above the blotting membrane(s) and a well on the flow distributor to contain the liquid to the desired area and to allow for lower starting volumes of such liquid. Preferably, the flow distributor is a non-binding or low binding hydrophilic porous membrane such as a 0.22 micron membrane.

The invention relates to the technical field of electro-analytical chemistry and protein identification transducers, and discloses a synthesizing method of functionalized ionic liquid and a preparation method and application of a molecular imprinting electrochemical transducer composed of the functionalized ionic liquid, carboxylation multiwalled carbon nanotubes and glassy carbon electrodes. The preparation method of the molecular imprinting electrochemical transducer comprises the steps that the polymerizable amino-functionalized ionic liquid is used as a functional monomer, BSA is used as template protein, N, N'-methylene bisacrylamide is used as a cross-linking agent, an oxidation-reduction system composed of ammonium persulfate and TEMED is used as an initiator, after polymerization, a molecularly imprinted polymer film is formed on the surfaces of the glassy carbon electrodes decorating the carboxylation multiwalled carbon nanotubes, and then the template protein is eluted to obtain the molecular imprinting electrochemical transducer which can identify template protein in a specific mode. The molecular imprinting electrochemical transducer has the advantages of being simple in preparation, low in material cost, high in selectivity, good in biocompatibility, and capable of being used for identifying and detecting protein in aqueous solution.

Provided are DNA microarray techniques that allow hybridization without RNA amplification, without using cDNA, and without labeling the nucleic acid prior to hybridization. Referred to as the Double-stranded Exonuclease Protection (DEP) assay, the technique permits the sample RNA to be used directly for hybridization, without manipulation in any way. Further provided is a microarray technique for high-throughput miRNA gene expression analyses, termed the RNA-primed, Array-based, Klenow Enzyme (RAKE) assay. The RAKE assay is a sensitive and specific technique for assessing single-stranded DNA and RNA targets, and offers specific advantages over Northern blots.

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.

Molecular species that are immobilized in discrete locations on a planar support such as protein bands on a gel or a blotting membrane or species applied in dots or spots on a membrane are reacted with binding reagents that are applied through a porous hydrophilic transfer sheet placed over the planar support, the sheet having at least one region that is laterally bordered by a barrier with the binding reagent retained within the bordered region. The bordered region is placed directly over an area on the planar support where the molecular species are expected to reside if they are present on the support. The binding reagent is then delivered into the support to contact the species. Targeted delivery of the binding reagent is thus achieved with improved efficiency.

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 discloses an up-conversion fluorescence immune chromatography test paper for the quantitative detection of neomycin and a preparation method thereof. The up-conversion fluorescence test paper comprises a support layer, an adsorption layer and a protection layer; the absorbing layer comprises an adsorption fiber layer, a fluorescent antibody fiber layer, a cellulose membrane layer and a water adsorption material layer at a handle hand; the cellulose membrane layer is provided with detection blotting printed by a carrier protein solution coupled with NEO and contrast blotting printed by a goat anti-mouse IgG; the fluorescent antibody adopts an NEO monoclonal antibody or polyclonal antibody marked by NaYF4:Yb:Er nanoparticles. Through the up-conversion fluorescence immune chromatography test paper for the quantitative detection of the neomycin, the application of the immune chromatography marked by up-conversion fluorescence nanometer materials in the quantitative detection of NEO residual is realized, so that the detection of the NEO residual is not subjected to background interference; the up-conversion fluorescence immune chromatography test paper for the quantitative detection of the neomycin is strong in specificity, high in sensitivity, simple, intuitional and accurate in detection, low in cost, wide in range of application and easy to popularize and apply.

The invention relates to a Raman spectrum detection method for a molecularly-imprinted membrane (MIM) by use of a Raman spectrometer used as a detection means with the combination of a molecular imprinting technology and a membrane technology and is used for rapidly detecting target molecules-dimethylguanidine hydrochloride and the like in real time. According to the method, a piece of lens paper for an optical lens is used as a substrate (carrier), guanidine hydrochloride or 4-(2-amino ethyl) benzsulfamide is used as an imprinted template, nanogold / silver sol is added in a pre-polymerization solution, and a polymer is directly light-gathered on the lens paper, so as to be convenient to carry and detect. The addition of the nanogold / silver sol is similar to the surface enhanced raman spectroscopy in principle, the response strength of the Raman spectrum can be enhanced and the fluorescence signal interference is reduced; the molecularly-imprinted membrane after and before target adsorption is compared with a standard substance, so that the peaked change in the Raman spectrum can be observed and whether a practical sample contains a target molecule can be judged.

The invention relates to a silk screen printing electrode and a method for synthesizing chloramphenicol molecularly imprinted membrane, belonging to the macromolecule technical field. The invention embeds template molecules in the direct surface in-situ polymerization of a one-time silk screen printing electrode and elutes the template molecules, to form the cavities of fixed sizes and shapes and the function groups of a fixed arrangement in the molecularly imprinted membrane, therefore, the molecularly imprinted membrane has shape memory property to the stereo structure of the template molecules, the chloramphenicol molecule of the object sample can be combined with the cavity specifically, to modify the electromechanical property of the molecularly imprinted membrane to improve the electric conductivity of the molecularly imprinted membrane and improve membrane passing current. The silk screen printing electrode coated with a chloramphenicol molecularly imprinted membrane can be connected with a current sensor to quickly and sensitively detect chloramphenicol density. The molecularly imprinted membrane can be used to detect chloramphenicol, having simple sample pretreatment, short detection time, strong interference resistance and batch production.

The invention discloses an immune chromatography test paper strip for the quantitative determination of clenbuterol based on up-conversion fluorescent nanoparticle label and a preparation method thereof. The test paper strip comprises a supporting layer, an adsorption layer, and a protection layer, wherein the adsorption layer comprises an adsorption fibrous layer, a fluorescent antibody fibrous layer, a cellulose film and an absorbent material layer at the handle end, the cellulose film is provided with detection blots printed by using a CL coupling carrier protein solution and control blots printed by using goat anti-mouse IgG antibodies, and the fluorescent antibody is an NaYF4:Yb:Er nanoparticle labelled CL monoclonal antibody or polyclonal antibody. According to the invention, the application of immune chromatography based on the up-conversion fluorescent nanoparticle label in the quantitative determination of CL residues is realized, so that the detection of the CL residues has no background interference; and the test paper strip 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 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.

The invention relates to a test strip for rapidly detecting residues and a preparation method of the test strip. The bottom layer of the test strip is used as a supporting layer; an intermediate adsorption layer is fixed on the supporting layer; an external protection layer is fixed on the adsorption layer; the adsorption layer comprises an adsorption fiber layer, a gold-labeled antibody fiber layer for adsorbing and coupling lincomycin and a cellulose membrane layer sequentially arranged on the testing sample end and further comprises a water absorbing material layer arranged on the handle end; a detection blot printed by a lincomycin carried protein solution and a control blot printed by a goat anti- or rabbit anti-mouse IgG antibody solution are arranged on the cellulose membrane layer. The test strip is high in detection specificity, high in sensitivity and simple and fast in operation; detection results are displayed visually, directly and accurately; a special apparatus and a professional are not in need in the detection; and the test strip has the following characteristics of low cost, low investment as well as easiness for promotion and application.

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 present invention provides a gene encoding a recombinant porcine circovirus type 2 Cap protein and application thereof. The present invention aims to obtain a soluble recombinant PCV2 Cap protein, and adopts a PCR method for fusion of MPG and NLS partially deleted Cap protein N-terminal gene to construct a new gene; and then the new gene is cloned into an E. coli expression vector pET28a to obtain a recombinant plasmid; the recombinant plasmid is transformed into E. coli BL21 to obtain recombinant engineering bacteria; the recombinant engineering bacteria is subjected to induced expression by IPTG to obtain the soluble recombinant PCV2 Cap protein. SDS-PAGE and Western-blot identification shows that most of the recombinant protein MrCap is present in the bacterial lysis supernatant and is soluble; and the recombinant protein MrCap has high immunogenicity and immunoreactivity, and lays foundation for the development of a PCV2 antibody detection kit and subunit vaccines.

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 present invention provides devices, apparatuses and methods for automated processing of biological samples. This invention provides automated devices and automated methods of sequentially treating a biological sample with processing fluids in more than one washing basin. In some embodiments, the invention provides automated devices and automated methods of western blot processing. In some embodiments, the invention provides automated devices and automated methods of Southern blot processing. In some embodiments, the invention provides automated devices and automated methods of northern blot processing. In some embodiments, the invention provides automated devices and automated methods of staining biomolecules on solid supports. In some embodiments, the invention provides automated devices and automated methods of nucleic acid separation and isolation.

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 present invention includes a method for detecting and isolating sugarcane proteins that interact with the HC-Pro and P1 proteins of SrMV and other proteins involved in gene silencing, particularly in sugarcane. The method uses a two hybrid assay with an HC-Pro, P1, or other silencing-related protein-containing bait protein and a prey protein containing a polypeptide encoded by a DNA molecule in a cDNA library. The method also includes identification of false positives through reverse two-hybrid assays and using in vitro techniques such as farwestern blots or pull down assays where plant physiological conditions may be replicated. Finally, interactions may be confirmed in planta. Some novel proteins used in and discovered using the these methods are also identified. Methods of using viral and plant proteins to regulate silencing in plants such as sugarcane are also discussed.

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 relates to the field of pesticide detection, in particular to a method and a kit for detecting atrazine, chlorpyrifos and triazolone based on an SERS technology. The kit is used for detecting one or more of the atrazine, the chlorpyrifos and the triazolone, and comprises a solid-phase carrier and labeled probes; the solid-phase carrier is coated with molecularly imprinted polymer membranes of the atrazine, the chlorpyrifos and the triazolone; the labeled probes are metal nanoparticles coupled with artificial antigens and labeled molecules; and the artificial antigen is one of artificial antigens of the atrazine, the chlorpyrifos and the triazolone, and the labeled molecules on the labeled probes can enable the atrazine, the chlorpyrifos and the triazolone to be distinguishedwhen the atrazine, the chlorpyrifos and the triazolone are detected by using the SERS technology. By combining a molecular imprinting technology with a Raman signal amplification probe technology, thedetection method for simultaneously detecting the three kinds of pesticides is established.

A preparation method of core-shell molecular imprinting nanometer silicon spheres comprises the following steps: synthesizing double bond-modified nanometer silicon spheres; synthesizing an auxiliary identification polymer chain with an identification group and an anchoring group which are randomly distributed; and self-assembling immunoglobulin heavy-chain binding protein BiP with the auxiliary identification polymer chain to prepare double-bond modified nanometer silicon spheres, and carrying out surface polymerization to form an imprinted polymer layer and an elution template. According to the invention, the prepared core-shell structured imprinting nanometer silicon spheres make imprinted sites be positioned on the material surface, substantially increase the imprinting specific surface area, and greatly increase the adsorption efficiency and adsorption quantity to target proteins; the existence of the auxiliary identification polymer chain makes imprinting have a specificity and a high efficiency, and the combination of the specificity and the high efficiency realizes the high-expansion and high-efficiency enrichment of natural microproteins and a strong practicality; and the use of cloned BiP as a template in the invention overcomes a problem that the natural microproteins cannot be obtained.