36 results about "Antibody labeling" patented technology

The invention discloses a culture method for inducing high expression of umbilical cord mesenchymal stem cells (LRP1) and application, and belongs to the technical field of biological medicines. The technical problem to be solved is to provide a standardized culture method for directionally inducing a specific functional phenotype cell population, so that efficient and accurate induction of LRP1 high-expression UCMSCs is realized, and clinical adaptation is met. According to the key point of the technical scheme, the culture method for inducing the LRP1 high-expression umbilical cord mesenchymal stem cells comprises the following steps: inoculating umbilical cord mesenchymal stem cells into a culture dish at the inoculation density of 5.5 * 10 < 3 > / cm < 2 > to 3.6 * 10 < 4 > / cm < 2 >, and culturing for 24-72 hours by using a special mesenchymal stem cell culture medium without adding a supplement, so that the acquisition efficiency of the LRP1 high-expression UCMSCs is improved, and the yield of the LRP1 high-expression UCMSCs is increased. Antibody labeling and flow sorting are not needed, so that the operation steps are simplified, and the complexity and cost of the culture process are reduced.

This invention discloses a quantitative test strip for HBV PreS1 antibody based on the AIE molecule, its preparation method, and its applications, relating to the fields of molecular biology, immunology, and biodetection technology. The test strip includes a sample pad, a labeling pad, a nitrocellulose membrane (NC membrane), an absorbent pad, and a base plate. The NC membrane detection line is coated with HBV PreS1 antigen to capture PreS1 antibodies in the sample. The labeling pad contains an immobilized Anti-Human IgG antibody conjugated to the AIE molecule, which binds to the PreS1 antibodies in the sample to form an immune complex. The test strip of this invention enables rapid detection of PreS1 antibodies and achieves enhanced fluorescence in the detection results. Combined with a fluorescence reader or fluorescence card reader, the detection results can be digitally and quantitatively analyzed. It is simple to operate, rapid in detection, and significantly improves detection sensitivity and enables quantitative analysis.

The invention relates to a method for detecting activation of specific T cells after tuberculosis infection and application of the method. Specifically, the invention provides a reagent combination for detection, and the reagent combination comprises an anti-CD4 antibody (CD4 antibody), an anti-CD154 antibody (CD154 antibody), an IFN-gamma detection antibody and a tuberculosis specific antigen. According to the method, through peripheral whole blood collection, PBMC cell separation, T cell activation (activation), activation termination (blocking), antibody labeling, membrane rupture, fixation, intracellular cytokine dyeing, flow cytometry detection and result analysis and judgment, the ratio of the expression IFN-gamma cells in the CD4 + CD154 + cells can be rapidly and accurately determined, and by combining an ROC curve and a constructed judgment standard, the content of the IFN-gamma cells in the CD4 + CD154 + cells can be determined, and the content of the IFN-gamma cells in the CD4 + CD154 + cells in the CD4 + CD154 + cells in the CD4 + CD154 + cells in the CD4 + CD154 + cells can be determined. And the sample to be detected is judged to be in a positive / negative / gray region, so that whether a patient is infected by the mycobacterium tuberculosis or not can be more accurately diagnosed, especially aiming at a whole blood sample from a special crowd.

The application discloses an acridinium ester labeled complex, which comprises a body, and a labeling group labeled on the body, wherein the body is an antibody, and the labeling group is an acridinium ester group. A preparation method of the acridinium ester labeled complex comprises the following steps: sucking a labeling buffer into a centrifugal tube; sucking an antibody to be labeled into the centrifugal tube and mixing to generate a first mixed solution; adding an acridinium ester solution and mixing, and then performing reaction at 25 DEG C in the dark for 2 hours to generate a second reaction solution; adding a termination buffer and mixing, and then performing reaction at 25 DEG C in the dark for 30 minutes to generate a third reaction solution; performing desalting purification on the third reaction solution to generate a fourth mixed solution; and filtering the fourth mixed solution with a 0.22 mu m filter membrane to generate the acridinium ester labeled complex. According to the preparation method, the labeling efficiency of the acridinium ester and the antibody is higher, the reactivity is stronger, the non-specific adsorption is lower, the sensitivity is higher, the repeatability and stability are better, and the labeling method is simple, rapid and easy to repeat.

The invention discloses a universal test strip for rapid detection of multiple nucleic acid amplification products as well as a preparation method and application of the universal test strip, and belongs to the technical field of rapid detection of nucleic acid. The universal test strip comprises a reaction bottom plate, a sample pad, a combination pad, a chromatography membrane and a water absorption pad, wherein the combination pad is loaded with a marked first target antibody, a marked first quality control antibody or antigen; the chromatography membrane is provided with a detection line T coated with a second target antibody and a quality control line C coated with a second quality control antibody or antigen; the detection line and the quality control line are sequentially arranged on the chromatography membrane in parallel at intervals along the chromatography direction. The universal test strip has the remarkable advantages of simplicity and convenience in operation, high detection speed, high sensitivity, strong specificity, low cost and the like, is wide in application technical range, is suitable for various nucleic acid detection technical methods, has strong tolerance to sample inhibitors, and is particularly suitable for being combined with isothermal amplification rapid detection technologies such as RPA (recombinase polymerase amplification), LAMP (loop-mediated isothermal amplification) and the like.

This invention relates to the field of biological detection, and more particularly to the preparation method and application of the t-PAI·C detection kit. The invention provides a detection reagent comprising: antibody-coated magnetic microparticles and enzyme-labeled antibodies; the antibodies include: tissue plasminogen activator-plasminogen activator inhibitor-1 complex antibody. In the detection reagent provided by this invention, when the sample contains the tissue plasminogen activator-plasminogen activator inhibitor-1 complex, the antibody coated with the magnetic microparticles and the enzyme-labeled antibody undergo an immune reaction with the complex, forming a sandwich-like immune complex consisting of the antibody coated with the magnetic microparticles, the complex, and the enzyme-labeled antibody; the enzyme on this complex acts on a luminescent substrate and emits light, and the concentration of the complex in the sample is calculated by detecting the luminescence signal.

The application discloses a pet inflammation multi-channel saliva detection test strip combined pad, a method and application, and particularly relates to the technical field of biomedical detection. The combined pad comprises a porous matrix, a polydopamine (PDA) bottom layer formed on the surface of the matrix, a poly(N,N-diethylaminoethyl methacrylate) (PDEAEMA) brush polymer layer grafted on the surface of the PDA bottom layer, and an antibody-labeled nanoparticle conjugate loaded in the polymer layer. The PDEAEMA brush polymer layer undergoes conformational change within the saliva pH range, shields and protects the antibody-labeled nanoparticle conjugate under higher pH conditions, and releases the antibody-labeled nanoparticle conjugate to participate in lateral chromatography under lower pH conditions. The multi-channel saliva detection test strip based on the combined pad forms multiple independent detection zones through a hydrophobic isolation structure, and is used for parallel detection of different inflammation markers.

The invention is applicable to the technical field of in-vitro diagnosis, and provides a centrifugal-lateral flow immune micro-fluidic chip for combined detection of allergens and a component sIgE, the centrifugal-lateral flow immune micro-fluidic chip comprises a chip body, and the chip body is fused with a centrifugal micro-fluidic functional area and a lateral flow micro-fluidic functional area; according to the method, an indirect immunoassay mode of firstly adding a specimen, capturing a to-be-detected antibody by an antigen probe to form a compound, capturing the compound by a microfluidic detection unit streptavidin, then dissolving a labeled antibody, combining the labeled antibody with the antigen probe and obtaining the to-be-detected antibody compound is adopted; the reaction is firstly completed in the reaction unit and then enters the lateral flow channel, at least two allergens / components can be synchronously detected, the sample dosage is less than or equal to 10 microliters, the detection time is less than or equal to 30 minutes, the sensitivity is improved by more than 10 times compared with that of a traditional method, and the method is adaptive to grassroots, emergency treatment and children screening scenes.

The invention relates to the technical field of biological detection, and discloses a salmonella monoclonal antibody, a salmonella colloidal gold immunochromatography test strip based on the antibody and a preparation method of the salmonella colloidal gold immunochromatography test strip. Salmonella is used as an antigen, a polyclonal antibody and a monoclonal antibody are prepared respectively, the monoclonal antibody 8E71C8 is screened out, and the salmonella colloidal gold immunochromatography test strip is prepared. The purified monoclonal antibody is used as a 30 nm colloidal gold marker, the optimal pH and the optimal antibody labeling amount of binding of the monoclonal antibody and colloidal gold and the concentration of the polyclonal antibody used as a detection line are optimized, the prepared salmonella colloidal gold immunochromatography test strip is high in specificity, the sensitivity is about 1 * 10 < 4 > CFU / mL and is higher than the average detection lower limit of domestic similar products, and the salmonella colloidal gold immunochromatography test strip can be used for detecting salmonella. And the test strip is simple to operate and high in efficiency, can be used as a means for rapidly and visually detecting the salmonella, and has popularization and application values.

The application discloses a kind of DOC-Chaps synergistic core's clinical and animal tissue rapid transparent reagent and three-dimensional imaging method. Rapid transparent reagent is mainly dissolved in deionized water by sodium deoxycholate (DOC), 3-[(3-cholamide propyl) dimethylamino]-1-propane sulfonic acid (Chaps), urea and 1-methylimidazole; DOC efficiently dissolves high-density lipid, Chaps gently destroys membrane structure, both complement each other, realize lipid rapid, depth clearance and significantly reduce light scattering; urea loosens collagen fiber and protein aggregate, improves the penetration rate of reagent; 1-methylimidazole selectively removes hemoglobin, reduces light absorption. The transparent reagent of the application can make millimeter thick brain slices in 4 (mouse brain) / 6 (human brain) hours The light transmittance of specific wavelength is greater than or equal to 90%, and there is no obvious tissue shrinkage / expansion, the compatibility of fluorescent protein and antibody label is excellent, suitable for three-dimensional imaging of high-lipid, high-hemoglobin samples such as in-vitro human brain, kidney, liver and mouse brain, heart, kidney, liver, intestinal tract, embryo, etc.

The invention provides an outer vesicle surface protein in-situ labeling method, and belongs to the technical field of outer vesicles, and the outer vesicle surface protein in-situ labeling method provided by the invention comprises the following steps: carrying out differential centrifugal separation on outer vesicles, carrying out quality evaluation by adopting nano-particle tracking analysis, and establishing a positive and negative correlation coupling equation set to optimize labeling conditions. The method comprises the following steps: fixing a sample by using glutaraldehyde, labeling antibodies in an optimized labeling buffer solution, removing free antibodies through ultrafiltration centrifugation, carrying out singular value decomposition calculation, adding a special protective agent, analyzing morphological characteristics by adopting a deep learning model, and ensuring the stability of the sample through filter membrane filtration and low-temperature preservation. In the whole process, a complete quality control system is established, the labeling targets of high specificity, high efficiency and high stability are achieved, and the technical problems of low labeling specificity and poor stability in the in-situ labeling process of the outer vesicle surface protein in the prior art are solved.

The invention provides a method for removing background fluorescence of a tissue sample, which comprises the following steps: mixing the tissue sample with a metal hydride reducing agent, and illuminating the tissue sample under white light and a light source with the wavelength of 475-485 nm. According to the method, the background fluorescence of the tissue sample can be obviously removed, the background is cleaner after the treated tissue is subjected to antibody dyeing, the strength and expression quantity of an antibody labeling signal are not influenced, the signal-to-noise ratio of a specific fluorescence labeling signal is high, the removal effect is thorough, and the background fluorescence cannot be recovered. Meanwhile, the method disclosed by the invention is low in cost and simple and convenient to operate, and has a wide application prospect in technologies such as immunofluorescence analysis and the like.

The invention discloses a large-volume sample fluorescence labeling method based on tissue fixation. The fluorescence labeling method comprises the following steps: a sample fixing step: fixing a biological tissue sample by adopting paraformaldehyde; and a fluorescence labeling step: placing the biological tissue sample subjected to fixation treatment in a stationary liquid containing glyoxal, and realizing fluorescence labeling of the biological tissue sample by utilizing autofluorescence generated by browning due to fixation of glyoxal. According to the fluorescent labeling method, fluorescent dye and antibody labeling are not needed, labeling of cell bodies and fiber bundles can be achieved in a non-degreasing state, and meanwhile blood vessel information can be obtained through a contrast means; moreover, the method is easy to process, the sample can be rapidly and uniformly permeated, the sample processing time is greatly shortened, and a new way is opened up for obtaining three-dimensional structure information of different large sample tissues and organs at the submicron scale.

This invention relates to the field of biological detection, and more particularly to antibody combinations and their application in the preparation of detection products for TPS. The monoclonal antibodies provided in this invention are named 7#, 13#, and 16#. This invention also discloses the preparation method and application of these antibodies. This invention uses prokaryotically recombinantly expressed CK18 as an immunogen to immunize mice. Sixteen anti-TPS monoclonal antibodies were obtained through screening and purification using hybridoma technology. The obtained antibodies were labeled with horseradish peroxidase. Epitope identification was performed on the synthesized peptides, and cross-pairing of different epitope antibodies was used to select one antibody suitable for coating and two antibodies suitable for labeling. These were then applied to detect TPS in human serum. The double-antibody sandwich detection method established using the above antibodies can be applied to a magnetic microparticle chemiluminescence platform, and has the advantages of high sensitivity, simple operation, and short detection time. It can provide key materials for the subsequent development of commercial kits for TPS detection.

The invention discloses an application of FXYD1 protein in sperm centromere detection. The amino acid sequence of the FXYD1 protein is as shown in SEQ ID NO. 1. By identifying an antibody of the FXYD1 protein, the expression position of the FXYD1 protein on the sperm is marked, and the number and position of centromere in the sperm are analyzed by combining immunostaining and microscope observation. The method can effectively evaluate the sperm centromere structure state, including existence, number and position form of centromere; the related expression of the sperm FXYD1 protein can also be evaluated. The human sperm quality evaluation method provided by the invention is helpful for objectively judging sperm functions, embryonic development after fertilization, physiological and pathological states related to male fertility and prognosis results of the physiological and pathological states, and has important transformation and popularization values and wide application prospects in the fields of reproductive biology and reproductive medicine.

The invention discloses a method for detecting quinoxaline antibiotics and main metabolites thereof, and relates to the field of agriculture and forestry chemistry detection.The method comprises the following steps that 1, a UCNPs-mAb fluorescent probe is prepared; step 2, preparing and assembling UCNPs-LFSBs; step 3, optimizing a UCNPs-LFSBs system, wherein the optimization content comprises the antibody labeling amount, the concentration of OLA-OVA, QCT-OVA and MQCA-BSA on a T line, the concentration of GAM-IgG on a C line, the mixing ratio of UCNPs-mAb mixed probes on a conjugate pad, the artificial antigen membrane scratching position, a buffer solution and the reaction time; by means of the optimized detection method, OLA, QCT and MQCA can be rapidly and quantitatively measured at the same time with high sensitivity.

The invention discloses a method for screening umbilical cord mesenchymal stem cells with an anti-depression function and a therapeutic drug. The method comprises the following steps: acquiring an umbilical cord mesenchymal stem cell sample; a CD146-PE antibody is used for marking sample cells, and flow cytometry is adopted for detection; determining the proportion of CD146 antibody-labeled positive cell subgroups in the sample; when the proportion is greater than 50%, the sample is judged to have an anti-depression function, and the effect of the sample in treating depression is predicted to be remarkable. The method can be used for efficiently screening the umbilical cord mesenchymal stem cell subpopulation for relieving depression, and has important medical application value and economic value.

The invention discloses a cat triple fluorescence immunochromatography antibody detection test strip as well as a preparation method and application thereof. The test strip comprises a bottom plate as well as a sample pad, a detection film and a water absorption pad which are sequentially connected on the bottom plate, a quality control microsphere line, a quality control line, a detection microsphere line, a first detection line, a second detection line and a third detection line are sequentially arranged on the detection film, the quality control microsphere line is coated with quality control antibody labeled microspheres, and the quality control antibody labeled microspheres are coated with quality control antibody labeled microspheres. The detection microsphere line is coated with antigen labeled microspheres, and antigens comprise a cat herpes antigen, a cat cup antigen and a cat plague antigen. The test strip realizes synchronous, rapid and accurate quantitative detection of the cat triple antibody, and has the characteristics of simplicity and convenience in operation, stable signal, good repeatability and wide linear range.

Provided is an antibody labeling method. The method comprises using a direct or indirect labeling method to directly couple a target label to an antibody. A flexible platform method is provided for a site-directed antibody labeling process, an antibody recombination process, and a multi-label coupling process.

The present application relates to a kind of inhaled allergen basophil activation detection method and its application.Specifically, the present application provides a reagent combination for detection, which includes: a first antibody against human CD203c, a second antibody against human CD123, a third antibody against human CD45, a fourth antibody against human CD63, a positive control, and an inhaled allergen.The method of the present application can quickly and accurately detect basophil activation by blood collection, activation reagent configuration, activation, termination of activation, antibody labeling, red blood cell lysis, flow cytometry detection, result analysis and judgment.The present application can be used for diagnosing inhaled allergen sensitized allergic diseases, identifying inhaled allergen and its components, predicting the severity of allergy and monitoring the efficacy.

The invention discloses preparation of ABEI macrolide and application of ABEI macrolide in preparation of an AD diagnostic kit, ABEI macrolide is adopted as a marker, the chemiluminescence reaction does not need to depend on horse radish peroxidase (HRP) and a traditional enhancer, and a detection system is simpler. The AD diagnostic kit provided by the invention comprises a reagent 1 containing magnetic beads labeled by a p-Tau181 antibody, a reagent 2 containing a p-Tau181 antibody labeled by ABEI macrolide, a p-Tau181 antigen standard substance and excitation liquid, and the provided AD diagnostic kit has the advantages of high stability, simplicity and convenience in operation, light drifting resistance, high sensitivity and low cost, and can provide more accurate detection guarantee for clinical diagnosis.

The invention belongs to the technical field of biological medicine. More specifically, the invention relates to a method for rapidly determining the virus replication period. Through high-frequency sampling after infection and combination of virus specific fluorescent antibody labeling and continuous observation in the same visual field, the invention provides a brand-new rapid determination method for the virus replication period. The method does not need to depend on an animal model and is completely completed in an in-vitro cell system, so that the operation cost is remarkably reduced; the shortest total experiment time can be controlled within 24 hours, and the research and development efficiency is greatly improved. The determination method provided by the invention has high specificity and sensitivity, is simple and convenient to operate, low in equipment threshold and suitable for various viruses, provides a brand new method for determining the virus replication period with speed, precision and feasibility for basic research of virology and research and development of antiviral products, and has good application prospects and application values.

The invention provides a nano-microsphere immunochromatography test strip for specifically detecting potato virus M. The test strip comprises a PVC (polyvinyl chloride) bottom plate, and a sample pad, a conjugate pad, an NC (nitrocellulose) membrane and a water absorption pad which are sequentially overlapped on the PVC bottom plate along a sample chromatography direction, a polystyrene nano-microsphere labeled antibody is fixed on the combination pad, a detection line T line and a quality control line C line are arranged on the NC membrane, a coated antibody is marked on the T line, and a goat anti-chicken antibody is marked on the C line; the labeled antibody and the coating antibody are paired monoclonal antibodies specifically combined with potato M virus coat protein PVM-CP; the labeled antibody is PVM-1G5B2F2 and is an IgG type antibody, the amino acid sequence of a heavy chain variable region is as shown in SEQ ID No.1, a light chain is a K type light chain, and the amino acid sequence of a light chain variable region is as shown in SEQ ID No.2. The test strip disclosed by the invention has the advantages of high sensitivity, high specificity, good stability, simplicity and rapidness in operation, high detection accuracy, wide application range and the like, and shows a good application prospect in rapid field detection of the potato virus M.

Engineered version of protein M, which binds to both κ and λ light chains and enables the labeling of all IgGs. The disclosed Engineered version of protein M enables the coupling of a wide range of molecules, such as luciferase domains, epitope tags, fluorescent domains, oligonucleotides, reporter enzymes, imaging tags, and small molecules to not only commonly used IgGs, but also chicken IgYs, Fabs, and scFvs, while still preserving effective antigen binding.

The invention discloses a clinical and animal tissue rapid transparentizing reagent taking DOC-Chaps cooperation as a core and a three-dimensional imaging method. The rapid transparentizing reagent is mainly prepared by dissolving sodium deoxycholate (DOC), 3-[(3-cholamidopropyl) dimethylamino]-1-propanesulfonic acid (Chaps), urea and 1-methylimidazole in deionized water; dOC efficiently dissolves high-density lipid, Chaps mildly destroys a membrane structure, and DOC and Chaps complement each other and cooperate with each other, so that rapid and deep removal of lipid is realized, and light scattering is remarkably reduced; urea loosens collagenous fibers and protein aggregates, so that the permeation rate of the reagent is increased; 1-methylimidazole is used for selectively removing heme and reducing light absorption. According to the transparent reagent, the light transmittance of a millimeter-thickness brain slice at a specific wavelength can be larger than or equal to 90% within 4 (mouse brain) / 6 (human brain) hours, no obvious tissue contraction / expansion exists, the compatibility of fluorescent protein and antibody labeling is excellent, and the transparent reagent is suitable for three-dimensional imaging of high-lipid and high-heme samples such as in-vitro human brain, kidney and liver and mouse brain, heart, kidney, liver, intestinal tract and embryo.

The invention discloses a method for quantifying and distinguishing MSC immunocompetence by using CD82 and application, and the method comprises the following steps: collecting cells, labeling membrane protein CD82 on the surface of MSC by using a fluorescently-labeled antibody, detecting the positive rate of the cell CD82 and MFI by using a flow cytometry, and quantifying the MSC immunocompetence through the expression level of the CD82. The immunocompetence refers to the inhibition capability of the MSC on inflammatory cytokines TNF-alpha and IFN-gamma secreted by activated PBMC, meanwhile, the MSC cell population with high immunocompetence can be distinguished by detecting the level of expression CD82 of the MSC, and the MSC cell population with high immunocompetence screened by the method shows a good curative effect in treatment of inflammation, immunity and other related diseases, and has good application prospects. The detection method is simple and easy to implement, easy to operate, economical and efficient, and has wide application prospects in the stem cell medicine market.

The application discloses a kind of for neuron intranuclear inclusion disease vascular sympathetic nerve injury evaluation method;Belong to the field of biological science and technology, its operating steps are as follows: skin biopsy, immunofluorescence staining, slide scanning, calculate vascular adrenergic small fiber density.The application obtains tissue by skin biopsy, is photographed after being given immunofluorescence staining, and the density of the sympathetic adrenergic small nerve of the superficial layer of dermis blood vessel marked by TH antibody is calculated using Image J software, to accurately assess whether the receptor of neuron intranuclear inclusion disease exists sympathetic nerve injury and its severity.

The application discloses a method for quantifying and distinguishing MSC immunological activity by CD82 and application, collects cells, uses a fluorescently labeled antibody to mark a membrane protein CD82 on a surface of MSC, and uses a flow cytometer to detect a positive rate and MFI of the cell CD82, quantifies the MSC immunological activity by an expression level of the CD82, the immunological activity refers to an inhibition ability of the MSC on secretion of inflammatory cytokines TNF-alpha and IFN-gamma by activated PBMC, meanwhile, the MSC cell group with high immunological activity can be distinguished by detecting the expression level of the CD82 on the MSC, the MSC cell group with high immunological activity screened by using the method shows good curative effect in treating diseases related to inflammation and immunity, the detection method is simple, easy to operate, economical and efficient, and has wide application prospect in a stem cell drug market.

This application relates to the field of in vitro bioassay technology, and discloses a method for preparing a colloidal gold assay reagent, its product, and its application. The method for preparing the colloidal gold assay reagent comprises five steps: colloidal gold preparation, antibody labeling, double blocking, centrifugal purification, and dilution and reconstitution. In particular, the double blocking process involves a first blocking agent selected from one or more of bovine serum albumin, ovalbumin, and casein; and a second blocking agent selected from one or more of lysine, glycine, alanine, serine, threonine, valine, and asparagine. This allows the prepared colloidal gold assay reagent to achieve high sensitivity and excellent C / T line color development when applied to FSH detection, reducing false positives caused by nonspecific adsorption and improving the accuracy of test results. Its limit of detection (LOD) is as low as 10 mIU / mL, solving the problems of low sensitivity and uneven C / T line color development in existing FSH assays.