50 results about "IMMUNE FLUORESCENCE" patented technology

The invention discloses an anti-human inflammatory factor interleukin 6 monoclonal antibody and an application thereof. When the monoclonal antibody is prepared, an adopted immunogen is a human inflammatory factor interleukin 6 antigen, and an amino acid sequence of the human inflammatory factor interleukin 6 antigen comprises a sequence as shown in SEQ ID NO: 1. A group of monoclonal antibodies specifically combined with antigens is constructed based on the IL-6 antigens. The IL-6 monoclonal antibody has strong binding capacity with an IL-6 antigen in a clinical sample, has no cross reaction with other proteins, has good specificity and affinity compared with a commercial antigen, can be effectively applied to ELISA and a dry-type immunofluorescence method, has good detection linearity, precision, stability and accuracy, and can be widely applied to the field of immunoassay. The antibody can be combined with the antigen to serve as a detection reagent raw material combination, and has clinical detection advantages.

The invention discloses a culture method of high-activity human urine-derived stem cells, which relates to the technical field of biological medicine, and is technically characterized by comprising the following steps: collecting urine samples of pregnant and lying-in women, coating a culture plate with gelatin, inducing adherence, performing passage amplification, performing immunofluorescence identification, performing function detection and other optimized conditions, and separating and culturing the stem cells. Compared with urine-derived stem cells of normal adults, the stem cells derived from pregnant women have stronger activity, higher dryness and faster multiplication capacity. The invention provides a novel and superior cell source for tissue engineering and regenerative medicine.

The application discloses a hybridoma cell strain 6F92E10, a monoclonal antibody thereof and application thereof, and belongs to the technical field of biotechnology.The application provides a hybridoma cell strain secreting an anti-mucinophilic Akkermansia monoclonal antibody.The antibody secreted by the hybridoma cell strain only combines with AKKermansia and does not cross-react with other common intestinal bacteria.In addition, the application solves how to detect the adhesion and colonization of mucinophilic Akkermansia in the colon of a mouse by using the prepared monoclonal antibody through an indirect immunofluorescence localization experiment, and expands the application range of the antibody.

The application provides a cell fixing solution for a myasthenia gravis antibody spectrum (IgG) detection kit (cellular immunofluorescence method) and a product and application thereof, and relates to the technical field of in-vitro detection. The cell fixing solution provided by the application is composed of polyoxymethylene, formaldehyde, acetic acid, raffinose and a buffer solution, can effectively fix the cell morphology, maximally retains the natural spatial conformation and antigen epitope integrity of myasthenia gravis related antigens AChR, Titin, RyR1, MuSK and LRP4, reduces the problems of antigen denaturation, epitope shielding and protein loss in the fixing process, can efficiently and accurately realize the synchronous joint detection of various specific antibodies, and the myasthenia gravis antibody spectrum (IgG) detection kit (cellular immunofluorescence method) prepared by the application has stable detection results, and has important clinical application value and industrial popularization prospect.

The invention discloses a rapid degreasing method for a biological tissue sample, which comprises the following steps: sequentially carrying out fixing, tissue dehydration, azeotropic degreasing, tissue rehydration and other treatments on the sample, and the degreasing solution is composed of alcohols and hydrocarbons according to a mass ratio of 1: 1-1: 2. According to the rapid degreasing method provided by the invention, the lipid in the tissue is efficiently extracted through mixed azeotropy of the alcohol and hydrocarbon organic reagents, so that the tissue is transparent and transparent; the problem of poor degreasing in tissues is solved by utilizing the high permeability characteristic of the micromolecular organic reagent; the dissolving capacity to phospholipid and cholesterol is improved by mixing organic reagents; through azeotropy, the degreasing efficiency is improved, the degreasing time is shortened, the internal permeability effect of the tissue is improved, and internal and external uniform permeability of the tissue is realized; by adjusting the components of the organic reagent, boiling rapid degreasing at low temperature is realized, and the retention of a fluorescence signal is improved; the tissue treated by the method is transparent and small in deformation, a fluorescent protein signal can be maintained, and the immunofluorescence staining effect can be improved.

The invention relates to the technical field of biological medicines, and discloses a blocking antibody targeting CXCL6 and an application of the blocking antibody in preparation of antitumor drugs. The antibody or antigen-binding fragment thereof is capable of binding specifically to human CXCL6 protein with high affinity (e.g., Kd < = 1 nM). According to the invention, the specificity and high affinity of the antibody are verified by Western Blot, immunofluorescence, immunohistochemistry and surface plasmon resonance (SPR) technologies. Functional experiments show that the antibody can effectively block CXCL6-induced neutrophil chemotaxis, including in Transwell, a three-dimensional gel model and a zebra fish living body model. Besides, in a mouse MC38 colon cancer model, the antibody shows remarkable anti-tumor activity and can inhibit tumor growth driven by CXCL6. The invention also provides a pharmaceutical composition containing the antibody, and application of the antibody in preparation of drugs for preventing or treating diseases (such as inflammatory diseases, autoimmune diseases and cancers) related to abnormal expression or activity of CXCL6.

The invention discloses application of ailanone in preparation of a medicine for treating feline infectious peritonitis, and belongs to the field of biological medicine. The change of the expression quantity of the feline infectious peritonitis virus N protein is detected through indirect immunofluorescence and western blot, and the change of the quantity titer of the feline infectious peritonitis virus is detected through TCID50. Results show that ailanone has an obvious effect of resisting the feline infectious peritonitis virus, the inhibitory effect on the feline infectious peritonitis virus is enhanced along with increase of the drug concentration, and the half effective concentration is 0.09 mu M. Meanwhile, drug inhibition tests show that ailanone has no inhibition effect on feline calicivirus. The invention provides a novel therapeutic drug and strategy for treating feline infectious peritonitis.

The application belongs to the technical field of immune detection, and particularly relates to a quantum dot labeled testosterone propionate direct competition fluorescence immune detection method. The application prepares a monoclonal antibody, and couples CdSe / ZnS (core / shell) quantum dots with the prepared anti-TP monoclonal antibody to synthesize a quantum dot (QDs) labeled anti-TP-mAb-QDs immune fluorescence probe, and construct a quantum dot labeled testosterone propionate direct competition fluorescence immune detection system for detecting testosterone propionate. The method for detecting testosterone propionate by the quantum dot labeled direct competition fluorescence immune detection method is simple in operation, and can detect the residual content of testosterone propionate in a sample to be detected without adding a chromogenic substance. The operation and reaction can be completed in one step. The method is simple, efficient, sensitive, low in detection cost, and can be applied to rapid detection of TP residues in feed, and has good accuracy.

The invention belongs to the technical field of biology, and particularly relates to a recombinant monoclonal antibody of sheep anti-bluetongue virus VP7 protein. A sheep anti-bluetongue virus antibody library is obtained through a single cell V (D) J sequencing technology, a sheep anti-bluetongue virus VP7 protein recombinant monoclonal antibody is screened and successfully expressed, and it is identified through indirect ELISA and cellular immunofluorescence experiments that the recombinant monoclonal antibody has high affinity with bluetongue virus and can be specifically combined with VP7 protein. The recombinant monoclonal antibody is used as a competitive antibody to establish a competitive ELISA detection method aiming at the bluetongue virus VP7 protein, high specificity of bluetongue antibody detection can be realized, and a basis is provided for mastering the epidemiology of bluetongue virus and evaluating the immune effect of vaccines.

This invention belongs to the field of biodetection technology, specifically relating to a material for detecting anti-Caspr1 antibodies and its preparation method. The invention constructs an expression plasmid capable of highly expressing the Caspr1 antigen, transfects it into HEK293T cells, and modifies the original antigen sequence during construction to achieve normal membrane expression. Subsequently, a modified cell culture protocol is used to obtain a material for detecting anti-Caspr1 antibodies based on cellular immunofluorescence assay. This invention, based on cellular immunofluorescence, avoids the limitations of flow cytometry, which is complex and requires expensive equipment. By modifying the protein expression mode to normal membrane expression, it avoids cell permeabilization, avoids false negatives caused by antigen conformational changes in enzyme-linked immunosorbent assay (ELISA) and Western blotting, and avoids false negatives caused by transmembrane expression due to transfection with the original Caspr1 sequence.

This invention belongs to the field of biodetection technology, specifically relating to a material for detecting anti-NF155 antibodies and its preparation method. This invention constructs an expression plasmid capable of highly expressing the NF155 antigen, transfects it into HEK293T cells, and modifies the pCDH vector with enhancers during construction. Subsequently, a modified cell culture protocol is used to obtain a material for detecting anti-NF155 antibodies based on a cell immunofluorescence methodology. This invention uses a cell immunofluorescence methodology to maximize the preservation of the antigen's spatial conformation through cell expression and antigen immobilization, preventing false negatives due to changes in binding sites. This invention selects the lentiviral vector pCDH to avoid the problem of non-specific binding that is difficult to distinguish, as the proteins expressed by common target gene vectors such as pcDNA3.1 may themselves contain antibody binding sites. This improves detection specificity and solves the problem of false negatives.

The invention relates to the technical field of antibodies, and particularly provides an anti-human TTC22v1 monoclonal antibody, a hybridoma cell strain prepared from the anti-human TTC22v1 monoclonal antibody and application of the anti-human TTC22v1 monoclonal antibody. The monoclonal antibody can specifically recognize the 432 -569 amino acid region of the TTC22v1 protein, and is secreted by a hybridoma cell strain TTC22v1-06 # with the preservation number of CCTCC (China Center For Type Culture Collection) NO: C202594. The antibody has high specificity and high sensitivity, can obviously distinguish TTC22v1 and TTC22v2 proteins, is especially suitable for Western blot and cell immunofluorescence detection, can effectively detect expression of endogenous TTC22v1 proteins in colon cancer cells, provides a reliable detection tool for research of TTC22v1 in tumor research and disease diagnosis, and has a wide application prospect.

The invention provides an anti-medetomidine monoclonal antibody as well as application and a product thereof, and relates to the technical field of biology. Medetomidine is used for immunizing a Balb / c mouse, splenocytes of the mouse are fused with myeloma cells, hybridoma cells with high specificity are obtained through specific high-throughput screening, a large amount of mouse ascites is obtained through culture and re-immunization, and the mouse ascites with high specificity is obtained. And the medetomidine monoclonal antibody with high purity, high sensitivity and high specificity is obtained through multi-step separation and purification, so that a required raw material is provided for developing an immune test strip for detecting medetomidine. The anti-medetomidine monoclonal antibody disclosed by the invention can be used for immunological detection such as immunoblotting and immunofluorescence, and the obtained antibody is proved to have good specific binding capacity.

The invention belongs to the technical field of biological medicines, and particularly relates to a human NSUN2 protein 190th lysine site lactic acid antigen, an antibody, a preparation method and application thereof. Relates to the technical field of lactylated antibody preparation, and the preparation method comprises the following steps: S01, carrying out lactylation on 190th lysine in a polypeptide of an amino acid sequence, and coupling with a carrier protein to obtain an initial antigen, the carrier protein comprising hemocyanin KLH; s02, injecting the obtained antigen into a living body culture sample for immune culture, and collecting whole blood after a preset period to obtain multi-antibody serum; and S03, purifying to obtain an iterative antibody. According to the invention, the expression difference in sensitive and drug-resistant tumor cells and tumor tissues can be detected by directly applying Western Blotting, immunofluorescence and immunohistochemistry, so that a powerful guarantee is provided for the detection of tumor sensitivity.

The application discloses a SjHNF4 polypeptide antibody, a preparation method and application. The preparation method comprises the following steps: downloading a SjHNF4 protein sequence, screening a polypeptide sequence, adding a cysteine at an N terminal, then performing polypeptide synthesis and purification, coupling to obtain a recombinant polypeptide, mixing the coupled polypeptide with Freund's complete adjuvant to obtain a mixture, immunizing a mammal by using the mixture, taking heart blood of the mammal and separating serum to obtain a polypeptide antibody. The polypeptide antibody is detected by using an ELISA method, and then the polypeptide antibody is purified by using a polypeptide affinity chromatographic column. Finally, the binding affinity of the antibody and the antigen is detected by Western Blot and immunofluorescence experiments. The polypeptide antibody obtained by the application has high purity (not less than 90%) and high titer (not less than 512000), and has good purity and immunogenicity, can satisfy various experimental requirements and has commercial value.

The invention discloses a hybridoma cell strain capable of secreting an anti-Ackermann muciniphila monoclonal antibody, the monoclonal antibody and application of the monoclonal antibody. Belongs to the technical field of biology. The invention provides a hybridoma cell strain capable of secreting an anti-Ackermann muciniphila monoclonal antibody. The secreted antibody is only combined with AKK bacteria and has no cross reaction with other common intestinal bacteria. Besides, through an indirect immunofluorescence localization experiment, the problem of how to detect the adhesion and colonization condition of Ackermann muciniphila in the colon of the mouse by using the prepared monoclonal antibody is solved, and the application range of the antibody is expanded.

The present invention discloses a shark single-domain antibody and its application that recognizes a unique specific B cell antigen epitope (LHRSYLTPGDSSSGWTAG) in the surface spike protein S of the new coronavirus (SARS-CoV2). The nucleotide sequence of the shark single-domain antibody is shown in SEQ ID NO: 1, and the amino acid sequence is shown in SEQ ID NO: 2. The shark single-domain antibody of the present invention has strong specificity, high sensitivity and good accuracy because it recognizes the unique B cell antigen epitope in the spike protein S of the new coronavirus (SARS-CoV2). It has been proven to be useful for detection such as enzyme-linked immunosorbent assay (ELISA), protein immunoblotting (WB) and cell immunofluorescence (IF).

This invention discloses an anti-ERBB2 monoclonal antibody, its preparation method, and its applications, belonging to the field of biomedical technology. This anti-ERBB2 monoclonal antibody uses the intracellular domain of the human ERBB2 protein as an immunogen and is obtained through animal immunization, B cell screening, gene cloning, and recombinant expression techniques. It can specifically recognize the human ERBB2 protein, exhibiting high affinity and high specificity, and can recognize both native and denatured ERBB2 proteins. This monoclonal antibody can be applied in immunohistochemistry, immunofluorescence, Western blotting (WB), flow cytometry, ELISA detection, and tumor targeted therapy. Experimental results show that this antibody has low background and no non-specific binding, accurately detecting ERBB2-overexpressing tumor cells and tissues, and has significant application value in the diagnosis, prognostic assessment, and targeted therapy of ERBB2-related diseases such as breast cancer.

The invention provides a rabbit polyclonal antibody aiming at primary cilia and a preparation method thereof, and belongs to the technical field of biology, and the preparation method comprises the following steps: taking a 200-428 amino acid sequence of human ARL13B protein as a target fragment, synthesizing a gene after codon optimization, cloning the gene to a pet-32a expression vector, and carrying out induced expression in escherichia coli to obtain the rabbit polyclonal antibody aiming at primary cilia. Purifying to obtain high-purity immunogen protein; the nucleotide sequence of the synthetic gene is as shown in SEQ ID NO.1, and the amino acid sequence of the high-purity immunogen protein is as shown in SEQ ID NO.2. Immunizing a New Zealand white rabbit with the high-purity immunogen protein to obtain immune serum; and purifying the immune serum to obtain the rabbit polyclonal antibody. The prepared rabbit polyclonal antibody has higher specificity and signal strength, the titer can reach 1: 64000 or above through ELISA detection, primary cilia can be marked more clearly in a paraffin section immunofluorescence experiment, background noise is remarkably reduced, and the reliability of an experiment result is improved.

The invention discloses a vagina micro-ecology automatic detection system and method based on immunofluorescence microscopic scanning and AI analysis. The system comprises a fluorescence imaging module, a motion control module, an image processing module and an AI intelligent analysis module. The motion control module is used for bearing a vaginal secretion slide sample and controlling the vaginal secretion slide sample to move in a three-dimensional space, and is matched with the fluorescence imaging module to carry out full-automatic scanning; the image processing module is used for enhancing and de-noising the collected original fluorescence image so as to improve the image quality; the AI analysis module performs multi-target recognition and quantitative analysis on the processed image based on a deep learning algorithm, and counts the number and proportion of various bacteria and cells; and finally, automatically generating a micro-ecology assessment report according to clinical rules. The full-flow automatic and intelligent detection of the vaginal secretion sample is realized, the problems of strong subjectivity and low efficiency of manual microscopic examination are effectively solved, and higher sensitivity, specificity and repeatability in vaginitis diagnosis are realized.

The application provides a kind of anti-hepatitis B HBcAg monoclonal antibody and its application and product, it is related to the field of biotechnology.The application uses HBcAg to immunize Balb / c mouse, takes mouse spleen cell and fuses with myeloma cell, and obtains hybridoma cell with high specificity by specific high-throughput screening, obtains a large number of mouse ascites by culture and re-immunization, and then obtains high-purity, high-sensitivity and high-specificity anti-HBcAg monoclonal antibody by multi-step separation and purification, which provides the required raw material for developing HBcAg detection immunostrip.The anti-HBcAg monoclonal antibody of the application can be used for immunological detection such as immunoblotting and immunofluorescence, and the obtained antibody has good specific binding capacity.

The present invention relates to the technical field of in vitro detection reagents, and specifically to a porcine circovirus antibody detection kit based on an indirect immunofluorescence method and its application. The porcine circovirus Cap antigen provided by the present invention can simultaneously bind to serum antibodies induced by two genotypes, TTSuV1 and TTSuV2, and has high specificity. Based on the Cap antigen and the IFA method, the present invention establishes a serological detection kit and detection method for TTSuV antibodies, which can simultaneously detect serum antibodies induced by two genotype viruses, TTSuV1 and TTSuV2, and has good specificity, stability and repeatability, effectively avoiding the problems encountered in the establishment process of the ELISA method, and can better meet the needs of TTSuV serological clinical detection, and has good application prospects.

The invention relates to the field of biological medicines, and discloses a targeted inhibitor capable of effectively inhibiting the progress of gallbladder cancer and remarkably improving the lifetime and prognosis of a patient. Through single-cell transcriptome data of a gallbladder cancer patient, single-cell transcriptome data of gallbladder cancer organoid, multiple immunofluorescence experiments, real-time quantitative PCR of gallbladder cancer cell lines, immunohistochemistry of gallbladder cancer tissues and organoid, gallbladder cancer whole exon data and inhibitor agonist experiments, it is found that compared with para-carcinoma, the single-cell transcriptome data of the gallbladder cancer patient, the single-cell transcriptome data of the gallbladder cancer organoid, the single-cell Differential genes of gallbladder cancer epithelial cells in cancer tissues are enriched to a WNT pathway, crosstalk is generated between the gallbladder cancer epithelial cells mainly through a WNT7A-FZD6 ligand and receptor pair in a WNT signal pathway, and inflammatory fibroblasts and epithelial cells can perform intercellular communication through a WNT5A-FZD6 ligand and receptor pair in an ncWNT signal pathway.

The application discloses an antibody of citrus psylla saliva protein DcMucin and a preparation method and application thereof. Through preparation of the DcMucin-like polyclonal antibody, it is verified for the first time that DcMucin-like is saliva protein of the citrus psylla, and is released into plant tissues to participate in saliva sheath formation with the citrus psylla feeding. The application fills the blank of the preparation and research of saliva protein antibodies of the citrus psylla, the prepared saliva protein polyclonal antibody of the citrus psylla has good specificity to the DcMucin recombinant protein, the antibody immunofluorescence labeling observation of the saliva sheath lays a foundation for further research on the biological function of DcMucin in the interaction of Huanglongbing bacteria-citrus psylla-citrus, and lays a foundation for further obtaining a specific target of green prevention and control of the citrus psylla.

This invention belongs to the field of biotechnology. It provides a recombinant protein whose amino acid sequence consists of two dominant epitopes of cardiac troponin I (cTnI) repeated in tandem. The amino acid sequence is converted into a corresponding nucleotide sequence using E. coli-preferred codons. The nucleotide sequence is then chemically synthesized and a recombinant expression vector is constructed, thereby increasing the expression level of the recombinant protein in E. coli. This invention also involves using this recombinant protein to immunize mice to establish a phage library, obtaining the corresponding cardiac troponin I single-chain antibody scfv sequence through panning and screening, constructing a complete mouse IgG1 antibody sequence expression vector from the obtained scfv sequence, expressing the monoclonal antibody via transient transfection of HEK293F cells, purifying the monoclonal antibody, and determining the optimal monoclonal antibody pairing using an immunofluorescence orthogonal experiment.

The invention belongs to the technical field of fluorescence detection, and particularly relates to a multi-target circulating tumor cell quality control kit as well as a preparation method and application thereof. The kit comprises multi-marker substance control microspheres, a fixed preservation system and a subpackaging unit, the microspheres are 2-6 [mu] m polymer substrate particles, the surfaces of the microspheres are covalently coupled with recombinant antigen proteins such as EpCAM, CK8, Vimentin and CD45, and different microsphere subgroups present multiple antigen combinations and are used for simulating marker characteristics of circulating tumor cells and immune cells. Through cross-linking fixation and freeze-drying protection treatment, the obtained quality control microspheres have good antigen stability and channel recognition performance, and are suitable for multi-platform quality control of a flow cytometer, a multi-channel immunofluorescence system and the like. The kit can replace traditional quality control cells, and standard, low-risk and long-term storage multi-target detection quality control is realized.

The invention discloses an overexpression antigen cell fixed dry-type preservation method for cellular immunofluorescence, particularly relates to the technical field of cell biology, and relates to an overexpression antigen cell fixed dry-type preservation method for cellular immunofluorescence. The overexpression antigen cell fixed dry-type preservation method comprises the following steps: cleaning to-be-preserved cells by using a 1 * PBS solution, fixing by using a PFA stationary liquid, cleaning again by using the 1 * PBS solution, repairing by using an antigen repairing liquid, sealing by using a sealing liquid, carrying out preservative treatment by using a preservation preservative liquid, and carrying out air-drying and cryopreservation. According to the present invention, the ammonium sulfate-based antigen repair liquid is introduced, the cells are subjected to mild treatment after fixation, and the antigen epitopes shielded due to paraformaldehyde excessive cross-linking are reversibly opened by using the suitable ion strength, such that the antigen and antibody binding capacity is significantly recovered, and the antigen binding preservation capacity of the overexpression antigen cells is improved.

The invention discloses a method for separating and culturing epididymal epithelial cells of cocks in vitro, and belongs to the technical field of cell separation and culture. The method comprises the following steps: aseptically collecting epididymis tissues of cocks of 20-25 weeks, performing impurity removal, cutting, combined enzyme digestion, gradient filtration and centrifugal cleaning, performing inoculated culture by using a special culture solution, and performing CK18 immunofluorescence identification after passage. According to the method for separation and in-vitro culture of the cock epididymal epithelial cells, the cock epididymal epithelial cell in-vitro model is established for the first time, the procedure is simple and convenient, the cost is low, impurity cells can be effectively removed, and in-vitro culture of the cock epididymal epithelial cells can avoid influences of in-vivo complex environments and individual differences; and a key cell model is provided for cock epididymis function research, sperm maturation and preservation and male sterility related research.

The invention discloses a hybridoma cell strain 6F92E10 as well as a monoclonal antibody and application of the hybridoma cell strain 6F92E10. Belongs to the technical field of biology. The invention provides a hybridoma cell strain capable of secreting an anti-Ackermann muciniphila monoclonal antibody. The secreted antibody is only combined with AKK bacteria and has no cross reaction with other common intestinal bacteria. Besides, through an indirect immunofluorescence localization experiment, the problem of how to detect the adhesion and colonization condition of Ackermann muciniphila in the colon of the mouse by using the prepared monoclonal antibody is solved, and the application range of the antibody is expanded.

The invention belongs to the technical field of biological detection, and particularly relates to a Claudin18.2 targeting nano antibody and application thereof. According to the present invention, the number of the provided nanometer antibodies capable of specifically binding Claudin18.2 is nine, the molecular weights of the nanometer antibodies are 12.9-14.9 kDa, the nanometer antibodies are respectively 2D11, 3A2, 3A5, 3C6, 3E6, 3H3, 4D4, 4F4 and 4G1, the amino acid sequences of the nanometer antibodies are respectively represented by SEQ ID NO.50-58, and the nucleotide sequences for coding the amino acid sequences are respectively represented by SEQ ID NO.59-67. The nano antibody provided by the invention is stable in structure and good in heat resistance, and can be used for preparing medicines for diagnosing and treating gastric cancer, pancreatic cancer, esophageal cancer, biliary tract cancer, colorectal cancer and lung cancer, and related products of the nano antibody can be used for preparing diagnostic reagents for immunodetection, flow detection and cellular immunofluorescence detection and medicines for treating diseases.