55 results about "Immunoblottings" patented technology

The invention belongs to the technical field of antibody preparation, and particularly relates to an anti-terminal deoxynucleotidyl transferase (TdT) antibody and application thereof. The amino acid sequences of CDR1-3 on a light chain variable region of the antibody are respectively shown as SEQ ID NO.3-5, and the amino acid sequences of CDR1-3 on a heavy chain variable region of the antibody are respectively shown as SEQ ID NO.8-10. The antibody disclosed by the invention can specifically recognize TdT protein naturally expressed in cells and tissues, has the advantages of high specificity, high affinity, high sensitivity and the like when being used for immunoblotting, immunohistochemistry and other immunodetection systems, is beneficial to reducing false positive and false negative rates in pathological diagnosis, improves the accuracy of immunodiagnosis, and has a good application prospect. The method has wide application potential in the field of immunological detection of living biological samples. In addition, the antibody disclosed by the invention has cross-species reaction characteristics on homologous TdT proteins of human, mice and rats, so that the application scene of the antibody is greatly expanded.

Disclosed herein are compositions comprising protein antigens and RNA encoding the same (eg., compositions comprising protein antigens and RNA encoding antigens) that can be used to induce an immune response against SARS-CoV-2. Also disclosed herein are immunogenic compositions and medical preparations comprising the same, and methods of making and using the same. In some embodiments, the technologies provided herein can be used to address and / or overcome immune imprinting in SARS-CoV-2.

The invention discloses a preparation method and application of carassius auratus rahn aquatic OmpA gene cloning and prokaryotic expression as well as a polyclonal antibody. The preparation method comprises the following steps: culture and DNA amplification of rahn aquatic, design of an OmpA primer, gene cloning and prokaryotic expression, back multi-point subcutaneous injection of an immune rabbit, heart blood sampling and preparation of the polyclonal antibody. The invention further provides OmpA protein prokaryotic expression and protein purification, preparation of OmpA polyclonal antiserum and application of fish bodies. According to the invention, a rahnella aquatilis OmpA gene is cloned, a specific Hind III and EcoR1 double-enzyme digestion primer sequence OmpA-1F / OmpA-1R is designed, a prokaryotic expression vector pET32a-OmpA is constructed, high-immunogenicity OmpA protein is obtained through purification, OmpA specific multi-antibody serum is prepared, and the OmpA specific multi-antibody serum can be used for preparing the multi-antibody serum. The OmpA polyclonal antibody can be used for detecting and positioning the expression application of rahnella aquatica OmpA protein in crucian carp bodies through bidirectional agar diffusion, an immunoblotting method and an immunohistochemical method, and provides a reference basis for research and development and application of rahnella aquatica subunit vaccines and a molecular diagnosis technology as well as immune prevention and control of infectious diseases of the rahnella aquatica subunit vaccines and the molecular diagnosis technology.

This invention discloses an anti-MSLN antibody and its applications. The anti-MSLN antibody comprises a heavy chain variable region and a light chain variable region. The heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3. The amino acid sequences of HCDR1, HCDR2, and HCDR3 are shown in SEQ ID NO:7, 8, and 9, respectively; and the amino acid sequences of LCDR1, LCDR2, and LCDR3 are shown in SEQ ID NO:4, 5, and 6, respectively. The antibody provided by this invention has good affinity and specificity for MSLN antigens, effectively detecting target proteins in cells overexpressing MSLN. It can also be widely used as a core ingredient in the development of high-sensitivity MSLN detection kits such as ELISA, Western blotting, and immunohistochemistry.

The present application relates to the antibody of phorbolylated isocitrate dehydrogenase 1 and its preparation method and purpose, and specifically provides a polyclonal antibody, the polyclonal antibody is obtained by immunizing animals with the antigen epitope peptide of IDH1 phorbolylated, the specific site of the antigen epitope peptide is phorbolylated, and high-titer antibodies can be prepared, the phorbolylated modification of IDH1 in clinical specimens can be recognized with high specificity, and the polyclonal antibody can be used for the research of cell / tissue specimen immunoblotting and immunohistochemical method and the diagnosis of IDH1 related diseases.

The invention provides a fish creatine kinase sensitization and immune cross reactivity analysis method which comprises the following steps: preparing a fish muscle water-soluble protein crude extract, separating 40-45kDa protein components in the fish muscle crude extract through SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis), and screening a target sensitization protein combined with IgE / IgG through immunoblotting; the method comprises the following steps: extracting total RNA of grass carp and synthesizing cDNA, designing a creatine kinase specific primer for amplification to obtain a creatine kinase gene sequence, and carrying out bioinformatics analysis after sequencing verification; the method comprises the following steps: cloning a creatine kinase gene to an expression vector, transforming escherichia coli BL21, performing IPTG induced expression, and performing urea gradient dialysis renaturation on an inclusion body to obtain the recombinant grass carp creatine kinase. Detecting the IgE binding activity of the recombinant grass carp creatine kinase and the serum of the allergic patient through dot hybridization; the IgG cross reactivity of the recombinant grass carp creatine kinase and the blue crab arginine kinase is verified through indirect ELISA; the IgE cross reaction degree of the recombined grass carp creatine kinase and the blue crab arginine kinase is quantified through inhibitory dot hybridization.

This invention discloses an anti-MSLN antibody and its applications. The anti-MSLN antibody comprises a heavy chain variable region and a light chain variable region. The heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3. The amino acid sequences of HCDR1, HCDR2, and HCDR3 are shown in SEQ ID NO:7, 8, and 9, respectively; and the amino acid sequences of LCDR1, LCDR2, and LCDR3 are shown in SEQ ID NO:4, 5, and 6, respectively. The antibody provided by this invention has good affinity and specificity for MSLN antigens, effectively detecting target proteins in cells overexpressing MSLN. It can also be widely used as a core ingredient in the development of high-sensitivity MSLN detection kits such as ELISA, Western blotting, and immunohistochemistry.

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.

The invention belongs to the technical field of antibody preparation, and particularly relates to an anti-MLH1 protein monoclonal antibody and application thereof in immunodetection. Amino acid sequences of CDR1-3 on a light chain variable region of the antibody are respectively shown as SEQ ID NO.3-5, and amino acid sequences of CDR1-3 on a heavy chain variable region of the antibody are respectively shown as SEQ ID NO.8-10. The antibody provided by the invention has strong specificity, high recognition sensitivity and good binding affinity for human MLH1 protein in cells and tissues, can accurately recognize and locate target protein in cells and tissues, greatly reduces the occurrence rate of false positive and false negative results, and can be applied to a plurality of immunodetection systems, such as human MLH1 protein, human MLH1 protein, human MLH1 protein, human MLH1 protein, human MLH1 protein and human MLH1 protein. Particularly, the method has good applicability in immunoblotting and immunohistochemical systems. Moreover, the antibody provided by the invention has cross reactivity to human and mouse homologous MLH1 proteins, and also has certain applicability and good application prospects in detection of mouse MLH1 proteins.

The invention provides a western blot image automatic analysis method, device, equipment and medium, and relates to the technical field of image understanding, the method comprises the following steps: obtaining a western blot image to be analyzed, converting the western blot image to be analyzed into a gray level image, and carrying out background noise reduction processing on the gray level image; generating a gray scale curve based on the gray scale values of the pixel points in the gray scale image; carrying out trough identification on the gray curve to obtain each trough point in the gray curve; and based on the position of the trough point in the gray curve, adding a segmentation line in the gray image to obtain a lane division result of the to-be-analyzed western blot image, and determining gray value data of each lane based on the lane division result. According to the method, lane automatic division and gray value extraction of the western blot image can be realized, and the analysis efficiency of the western blot image is improved.

The invention provides an anti-dsRNA antibody, and also provides a preparation method and application of the antibody. According to the invention, dsRNA antigen is adopted to immunize mice, and an anti-dsRNA monoclonal antibody is obtained through immune screening. The anti-dsRNA antibody disclosed by the invention is specifically combined with dsRNA, not only can be combined with unmodified dsRNA, but also can be combined with dsRNA modified by pUTP, N1-Me-pUTP and 5-OMe-UTP, and can be applied to detection of dsRNA by an immunoblotting method, an enzymatic immunoblotting method and an ELISA (Enzyme-Linked Immunosorbent Assay) method.

The invention belongs to the technical field of medicines, and particularly relates to an experimental method for treating liver cancer by using oleanolic acid. Comprising an in-vitro experiment method and an in-vivo experiment method. The in-vitro experiment method comprises the following steps: S1, cell culture; s2, cell proliferation and migration; s3, detecting cell apoptosis and a cell cycle; s4, performing immunoblotting; s5, carrying out real-time quantitative PCR; the in-vivo experiment method comprises the following steps: step 1, performing chick embryo chorioallantoic model experiment; and 2, carrying out tumor-bearing experiment, treatment and tissue sampling on the mouse. The application of the oleanolic acid in liver cancer treatment is systematically researched for the first time, it is found that the oleanolic acid has the remarkable effects of inhibiting proliferation, inducing apoptosis, resisting angiogenesis and the like on liver tumors through the synergistic effect of multiple mechanisms, the remarkable curative effect on cancer mice is achieved, and a new medicine choice is provided for liver cancer treatment.

This invention relates to a method for constructing a polycystic kidney disease (PCD) model and its applications. The method involves overexpressing the MYCN gene in a target animal to obtain a PCD model that leads to PCD-related phenotypes. The method includes the following steps: obtaining a first strain of mice with Rosa26 knock-in overexpressing the CAG-LSL-HA tag-MYCN-IRES-BFP-Wpre-polyA gene; obtaining a second strain of mice by inserting Cre-WPRE-polyA into the start codon of the Pax8 gene; and crossing the first and second strains of mice to obtain a MYCN-overexpressing PCD model. This invention employs various experimental methods for validation, including histopathological analysis, immunohistochemical staining, and Western blotting. The model provided by this invention overcomes the limitations of existing in vitro cell models, organoids, and existing animal models in terms of limited phenotypes. The established MYCN-overexpressing PCD animal model exhibits stable disease progression and a short disease cycle, which not only helps to elucidate the disease mechanism but also serves as an ideal platform for drug screening and efficacy evaluation.

This application discloses the use of ritonavir or its derivatives in the preparation of medicaments for treating mental disorders. This application also discloses the discovery of mice (AtLAS) with difficulty in extinction of fear memories. ‑ / ‑ Intraperitoneal injection of ritonavir significantly restored the rigidity rate in model mice. Further immunoblotting and co-precipitation confirmed that ritonavir could significantly block the binding of Syn2a and Syngr-3, and promote the formation of VAMP2, Syntaxin, and SNAP25 trimers, thereby promoting the normal release of presynaptic membrane vesicles in neurons and restoring the rigidity rate in mice with refractory rigidity, thus achieving the treatment of mental disorders such as post-traumatic stress disorder.

The application discloses an antibody (YHWG-1) against HLA-G molecules and application, the antibody (YHWG-1) is produced by hybridoma with the preservation number of CCTCC NO:202120, using all currently known 7 HLA-G isomer molecules (HLA-G1, HLA-G2, HLA-G3, HLA-G4, HLA-G5, HLA-G6 and HLA-G7) shared, the antigen peptide (QTDRMNLQTLRGYYNQSEAS) of the 72~91 amino acid sequence in the HLA-G molecule heavy chain alpha 1 domain is immunogen.The application provides nucleotide and the amino acid sequence coded by the YHWG-1 antibody of the application, and the application also provides the application of the YHWG-1 antibody for HLA-G isomer molecule immunohistochemical, immunoblotting and flow cytometry detection.

The utility model discloses an incubation device of an immunoblotting instrument. The incubation device comprises an incubation disc fixing frame, a heating plate, a rocker arm assembly, a synchronous moving assembly, a driving motor and a control circuit. The rocker arm assembly is connected with the incubation disc fixing frame through a hinge and used for driving the incubation disc fixing frame to shake, and synchronous moving assemblies are further installed on the two sides of the incubation disc frame and used for stabilizing the incubation disc fixing frame and shaking with the rocker arm assembly at the same frequency under driving of the rocker arm assembly. The device has the advantages of accurate temperature control, uniform shaking, simplicity and convenience in operation and the like, and the efficiency and the accuracy of an immunoblotting experiment are improved.

The invention provides an anti-sT2 monoclonal antibody as well as application and a product thereof, and relates to the technical field of biology. According to the invention, a Balb / C mouse and a New Zealand white rabbit are respectively immunized by using sT2 immunogen expressed and purified by a eukaryotic system, and the anti-sT2 monoclonal antibodies anti-sT2-mab1 and anti-sT2-mab2, which are high in purity, high in sensitivity, high in specificity and capable of recognizing different antigen epitopes, are obtained through multi-step screening by using a mouse hybridoma and rabbit B cell sorting method. Compared with the traditional homologous antibody pairing which is easy to cause signal weakening due to epitope steric hindrance, the anti-sT2-mab1 and anti-sT2-mab2 are combined, so that the species cross reaction can be obviously reduced. The antibody developed by the invention can be used for immunological detection of immunoblotting, immunofluorescence and the like.

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 discloses a method for extracting high-purity liver cell membranes of pelteobagrus fulvidraco. The method comprises the following steps: firstly, cutting the liver into pieces to obtain large liver tissues; digesting the large liver tissue for multiple times by using isopyknic protease liquid; adding an extraction reagent 1 containing a protease inhibitor and a phosphatase inhibitor into the digested small tissues and cell precipitates, and ultrasonically crushing to obtain homogenate; performing low-speed centrifugation on the homogenate for multiple times to collect supernate, and performing high-speed centrifugation for multiple times to collect clear liquid; carrying out ultracentrifugation to collect the precipitate, and resuspending; obtaining a coarse cell membrane suspension; adding the coarse cell membrane suspension into a centrifugal tube, then adding a sucrose density gradient buffer solution until the centrifugal tube is filled with the coarse cell membrane suspension, and carrying out ultracentrifugation; and resuspending the cell membrane precipitate to obtain the pelteobagrus fulvidraco liver cell membrane. Immunoblotting verification on the cell membrane and other organelles extracted by the method shows that the cell membrane extracted by the method conforms to the expected effect of high purity, and other organelles contained in the cell membrane have few impurities.

The invention relates to a method and a device for enhancing immunoblotting and immunoblotting electrophoresis sensitivity. The method comprises the following steps: performing regular stripe standing wave concentration on protein by utilizing regular stripe standing waves generated by superposing surface sound source ultrasonic waves through a gel electrophoresis carrier which is subjected to electrophoretic separation and contains a low-concentration protein molecule band in a direction vertical to electrophoretic separation; and the protein molecules with different densities are pushed to the node or antinode region of the standing wave and are concentrated. The device comprises an area sound source generator operated by the method and an echo carrier plate with an echo plate. An electrophoresis tank for accommodating an echo carrying disc is additionally arranged, so that the electrophoretic separation step and the concentration step are integrated; and a main bearing body is arranged for bearing the electrophoresis tank, a related power supply and a related switch. According to the method and the device, the concentration of the low-abundance protein in the reaction area reaches or is closer to the optimal concentration for antigen-antibody reaction, in the immunoblotting process, the local luminescence or fluorescence peak intensity after membrane transfer meets the condition of reaching a threshold value or above, and the detection rate of the low-abundance target protein is greatly improved.

The methods, compositions and kits described herein provide signal amplification approaches for the detection of target biomolecules that significantly increase the sensitivity of detection using target-binding ligand molecules. The methods include cyclic addition of nucleic acid repeats to, e.g., target-binding molecules in situ, thereby providing multiple landing pads for labeled probes. The methods are well-suited to performance in multiplex, thereby permitting the sensitive detection of multiple targets in a single assay. These methods and compositions can be applied to, among other things, imaging, flow cytometry, CyTOF, and immunoblotting, providing additional tools for research and diagnostic purposes.

This invention relates to the field of molecular biology, specifically to the application of PRAS40 protein as a therapeutic target for Alzheimer's disease. This invention combines Western blotting, computer-generated virtual docking, and surface plasmon resonance (SPR) techniques to confirm that PRAS40 protein is a key target influencing the pathological progression of AD. Experimental results show that specific intervention (elimination or reduction) of PRAS40 protein levels is sufficient to significantly improve cognitive deficits and pathological characteristics in AD model mice, providing a clear molecular target for subsequent screening and development of novel anti-AD drugs.

The application discloses an ELISA kit for detecting anti-human laminin gamma 1 IgA autoantibody and application thereof. The kit comprises an enzyme-labeled plate coated with human laminin gamma 1 recombinant protein. Preferably, the human laminin gamma 1 recombinant protein is deglycosylated human laminin gamma 1 recombinant protein. Further, the application also provides application of the kit in preparation of a reagent for diagnosing mucous membrane pemphigoid, in particular Laminin gamma 1 IgA type pemphigoid. The kit is the first kit for detecting human laminin gamma 1 IgA antibody by using the ELISA method, has the advantages of simple operation, short time consumption and low cost compared with traditional immunoblotting method, and adopts full-length deglycosylated laminin gamma 1 recombinant protein as an antigen, thereby solving the problem of incomplete antigen epitope coverage in the prior art, and early detection of diseases can be realized, early treatment is achieved, and the life quality of patients is improved.

The invention provides a qualitative immunoblot-based in vitro method for the detection of onconeural antibodies class IgG to twelve different antigens (amphiphysin, CV2, PNMa2 / Ta, Ri, Yo, Hu, recoverin, SOX1, titin, zic4, GAD65 and Tr (DNER)) in serum or plasma samples of mammalian animals such as dogs, cats, ferrets, and rabbits for early diagnosis of twenty two cancer and cancer-associated neurological diseases. Detection of these antibodies in the blood of the animals is confirmed via an indirect immunofluorescent assay. Examples, including enzyme anti-dog, ant-cat, anti-ferret, and anti-rabbit conjugates, and serum or plasma quantities, are provided.

This invention discloses a shark-derived nanobody targeting the SARS-CoV-2 RBD protein and its applications. The nanobody's nucleotide sequence is shown in SEQ ID NO: 1, and its amino acid sequence is shown in SEQ ID NO: 5. Preparation involves immunizing a striped bamboo shark with the RBD protein, extracting total RNA from peripheral blood lymphocytes and reverse transcribing it into cDNA. The amplified VNAR fragment is then seamlessly ligated with amplified pR2 phage particles, transformed into TG1 competent cells, and a phage antibody library is constructed. SARS-CoV-2 RBD protein-specific nanobody sequences are screened from the library and recombinantly expressed to obtain the targeted shark-derived nanobody. The nanobody of this invention has a small molecular weight, allowing for the detection of the SARS-CoV-2 RBD antigen using paired nanobodies. It can be applied to immunoblotting, enzyme-linked immunosorbent assays (ELISA), and COVID-19 pharmaceutical applications related to the SARS-CoV-2 RBD protein.

The application discloses a kind of for preventing or treating post myocardial infarction ventricular arrhythmia pharmaceutical composition, the pharmaceutical composition is by everolimus, ethmolam and fluoxetine, belongs to the field of medicine technology.Immunoblotting proves that everolimus, ethmolam and fluoxetine can inhibit fatty acid oxidation protein.In vitro and in vitro cardiac electrophysiology experiment proves that the pharmaceutical composition can effectively reduce post myocardial infarction ventricular arrhythmia susceptibility.TTC staining proves that the pharmaceutical composition can effectively reduce post myocardial infarction myocardial damage area.Based on the above composition in the regulation of post myocardial infarction ventricular arrhythmia susceptibility, it can be further developed in the clinical application of the disease.

The invention belongs to the technical field of biological medicines, and particularly discloses an application of cumin in the aspect of neuroinflammation resistance, which is specifically as follows: lipopolysaccharide with the final concentration of 1 mu g / mL is used for inducing BV2 cells to form an acute neuroinflammation model, and cumin with gradient concentration is respectively added for treatment. Results show that 50-200 [mu] M cumin can significantly down-regulate expression of inflammatory factor nitric oxide, tumor necrosis factor-alpha and interleukin-6 secreted by lipopolysaccharide-induced BV2 cells. An immunoblotting method is used for discovering that cumin aldehyde can reduce expression of neuroinflammation 15-lipoxygenase / inflammasome signal channel targets in a dose-dependent manner, and meanwhile inflammatory reaction can be relieved by activating an oxidative stress signal channel. The results show that cumin has an anti-neuroinflammation effect. The cumin has a good application prospect for developing drugs or anti-inflammatory health-care products, cosmetics and food additives for treating inflammatory diseases by using cumin.

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 invention relates to the technical field of biology, and particularly discloses a pea active peptide and application thereof in inhibiting porcine coronavirus. The toxicity of pea active peptides EHYDSEAILFKK with different concentrations on Vero cells and ST cells is determined through a CCK-8 test, and it is proved that the pea peptide has no toxic effect on the cells; indirect immunofluorescence, virus copy number, immunoblotting and half tissue infection amount are used for determining that the pea active peptide EHYDSEAILFKK has an inhibition effect on porcine epidemic diarrhea virus in Vero cells and porcine transmissible gastroenteritis virus in ST cells; animal experiments show that piglets with 200 [mu] M of pea active peptide can significantly inhibit in-vivo replication of PEDV and TGEV. Therefore, the pea active peptide can obviously inhibit the replication of the porcine epidemic diarrhea virus and the porcine transmissible gastroenteritis virus in vivo and in vitro, and has a broad-spectrum effect of resisting the porcine coronavirus.

The invention relates to the technical field of biological detection, and discloses a GtaA antigen based on a glycosylation modification form and application of the GtaA antigen in aspergillus fumigatus infection detection. The GtaA antigen is exosome glycoprotein from aspergillus, and the amino acid sequence of the GtaA antigen is as shown in SEQ ID NO: 2. The GtaA antigen comprises GtaA which is subjected to full glycosylation, GtaA (N) which is subjected to deglycosylation modification, and GtaA (N-O) which is subjected to non-glycosylation modification. The invention establishes an immunological detection method based on different forms of GtaA antigens. Experiments show that glycosylation modification affects antigen diagnosis performance, but GtaA in glycosylation and non-glycosylation forms can be used as specific diagnosis antigens of aspergillus infection. The invention is widely applicable to rapid detection of aspergillus fungal infection, including immunological detection modes such as enzyme-linked immunosorbent assay, immunochromatography, immunoblotting, chemiluminescence and the like, and has important clinical application potential and popularization value.