204 results about "Immunoblottings" patented technology

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

Disclosed are methods and apparatus for separation of biomolecules via two-dimensional gel electrophoresis, methods and apparatus for immunoblotting separated biomolecules, and methods for the use of biomolecules processed via the methods and apparatus of the present invention, including use in a clinical setting. The methods and apparatus for separation of biomolecules via two-dimensional gel comprises vertical agarose gel electrophoresis in the first dimension, and the electrophoresis of a novel non-denaturing 3-35% concave gradient polyacrylamide gel in the second dimension. This novel gel can be cast in a modified gel caster that can facilitate the pouring of multiple gels simultaneously. The methods and apparatus for immunblotting are useful with any type of immunoblotting, including Western blot, Northern blot, and Southern blot analyses. These methods and apparatus provide safe, efficient and cost-effective immunoblots, while facilitating the reduction of exposure to toxic or radioactive materials, as well as the disposal of those materials.

The invention relates to a method for preparing a genetic engineering product, in particular to a prokaryotic expression protein of VP73 gene from African swine fever virus (ASFV) and a preparation method thereof. The preparation method comprises: artificially synthesizing the whole-length sequence of VP73 gene according to the sequence of the VP73 gene from ASFV in GenBank, constructing a recombinant expression vector pET32a-VP73, sequencing, verifying, transforming prokaryotic expression recipient bacteria E.coli BL21(DE3), and inducing expression by isopropyl-1-thio-beta-d-galactopyranoside (IPTG), wherein the molecular weight of the recombinant fusion protein is about 65KD. Protein purified by nickel column affinity chromatography can undergo a specific immune imprinting reaction with ASFV positive serum and avoid cross reaction with viruses such as swine fever virus, hog cholera virus, porcine circovirus, porcine reproductive and respiratory syndrome virus, swine influenza virus and pseudorabiesvirus. Experiments show the expressed protein has high detection sensitivity, and high specificity. When the antigen is used for detection, risk of spreading poison is avoided. And the antigen can be used as a detection antigen for use in an enzyme-linked immuno sorbent assay (ELISA) method for identifying an ASFV antibody.

The invention aims at providing chimeric protein of enterovirus EV71 neutralization epitope and a human norovirus P structure domain and an establishing method of a chimeric vector. By means of immunoblotting, it is verified that the chimeric protein of the enterovirus EV71 neutralization epitope and the norovirus P structure domain can be used for research and development of detection kits of EV71 and norovirus and novel bivalent subunit vaccines.

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

The invention relates to an extraction method of protein from animal tissues, in particular to an extraction method of total protein from a rumen epithelial tissue of a milk cow. The method comprises the steps of putting a rumen epithelial tissue sample of the milk cow in a precooled mortar, adding liquid nitrogen, grinding into powder, adding an acetone solution of precooled trichloroacetic acid, oscillating, eddying, uniformly mixing for overnight aging, centrifugally collecting sediment, freeze-drying the tissue sediment, adding lysate, incubating and eddying at intervals, and centrifugally collecting supernate, namely the total protein of the rumen epithelial tissue of the milk cow. For the total protein of the rumen epithelial tissue of the milk cow, extracted by the method, a total protein mixture can be separated by a two-dimensional gel electrophoresis technology according to an isoelectric point and molecular weight of the protein, the relative abundance of the protein is acquired, the isoelectric point, the molecular weight and a relative transcript level of each protein can be clearly and visually shown in a gel atlas, and the method has the advantages of more obvious integrity and visualization in comparison with the prior arts such as a high efficiency liquid chromatography separation method, an enzyme-linked immunosorbent assay and immunoblotting.

The invention provides an establishing method of a Chinese tongue squamous cancer cell line (CTSC-1), and provides a material base for researching the characteristics of tongue squamous cancer and therapeutics thereof. The establishing method comprises the steps of: carrying out primary culture on the tongue squamous cancer cells by adopting a tissue explants adherent method; subculturing when the cell density reaches about 80 percent; carrying out RT-PCR (Reverse Transcription-Polymerase Chain Reaction) detection on stably subcultured cells to indicate that the cells express tongue squamous cancer cell symbolic molecules CK 8, 18 and 19; and detecting adherent cells through a western blot test to express proteins of the tongue squamous cancer cell symbolic molecules CK 8 and 18, therefore, proving on a molecular level the successful establishment of the CTSC-1.

The invention discloses a theileria equi immunoblotting detecting method and a method for preparing a kit, wherein the theileria equi immunoblotting detecting method in the invention is established on the basis of acquiring diagnostic antigen and positive control serum via Cochlearia officinalis hyperoxide enzyme label and chemical light emitting prime. The solvent for the method in the inventionis little in amount, low in cost, high in specificity, high in sensitivity, high and fast in operation, free of toxicity and harm, low in required conditions, easy for judging the test result, long-time for storage, and stronger in specificity than ELISA. The method in the invention detects no false positive and false negative result.

The invention discloses a method for detecting autoantibodies of a systemic lupus erythematosus patient. The method includes steps of: detecting antinuclear antibodies and double-strand deoxyribonucleic acid (DNA) of the systemic lupus erythematosus patient respectively in an indirect immunofluorescence method, measuring antinuclear antibodies spectrums in a euroimmun western blotting method, and finally detecting single-strand DNA of the systemic lupus erythematosus patient through an enzyme-linked immunosorbent assay. The method for detecting autoantibodies of the systemic lupus erythematosus patient simplifies the existing detection method, improves detection efficiency and accurate diagnosis rate, and is beneficial to clinical diagnosis and treatment of systemic lupus erythematosus.

The invention belongs to the field of life science research, and specifically relates to a method for detecting the expression quantity of nicotine acetylcholine receptors (nAChRs) in peripheral bloodlymphocytes of rats, which is characterized by comprising the following steps: 1) separating peripheral blood lymphocytes of rats; 2) acquiring cell total protein, concentration determination and sample preparation; and 3) detecting the expression quantity of the nAChRs in the lymphocytes by a western blot (WB) experiment. According to the invention, the detection of the protein expression in theanimal peripheral blood in real time can be realized by determining the protein in the peripheral blood lymphocytes, and the kit has important significance for the research of nicotine addiction biomarkers.

The invention provides a reaction membrane strip for detecting autoimmune diabetes, a preparation method and a using method, and belongs to the field of biomedical science. Aiming at the lack of an immunoblotting product special for various immune diabetes experimental examinations and physical examination at present and some defects of an existing immunoblotting agent, the invention provides the reaction membrane strip for detecting the autoimmune diabetes. The reaction membrane strip comprises a bottom lining plate, not less than one detecting zone and not less than one indicating zone, wherein the detecting zones and the indicating zones are arranged on the bottom lining plate; the detecting zones, the indicating zones as well as the detecting zones and the indicating zones are parallel to each other. Compared with the prior art, the reaction membrane strip provided by the invention has the following advantages that various autoantibodies can be detected through one clinic sample and once detection reaction, insufficient sensitivity and specificity of single autoantibody detection and operation complexity of independently detecting autoantibodies related to various diseases one by one are overcome, and the detecting efficiency and the result judgment accuracy are greatly increased.

The invention belongs to the technical field of immunoassay and in particular relates to a method for detecting trace protein by fluorescent immunoblotting of quantum dot nanospheres. The method comprises the following steps: firstly, carrying out gel electrophoresis separation on a protein mixture to be detected; transferring gel subjected to the electrophoresis separation into a solid-phase carrier film; then sealing a protein-loaded solid-phase film; after adding a specific primary antibody of protein to be detected, washing; then reacting with a fluorescent secondary antibody of the quantum dot nanosphere marker and washing; finally, developing and photographing through a protein blot film under the illumination of exciting light. Compared with chemiluminescence, the method for detecting the protein through the fluorescent immunoblotting, provided by the invention, has the characteristics that luminescent substrates are not needed, a detection signal is more stable and can be stored for a long period, results have consistency and repeatability and the like; compared with near-infrared fluorescent immunoblotting, the method has the characteristics that an additional signal amplification step is not needed, a fluorescent signal is more stable, light-shielding operation is not needed, an expensive near-infrared laser scanner does not need to be used for detecting and the like.

The invention relates to polypeptide biological technical field, which discloses a spatial conformation mimic antigen epitope of H3-type influenza virus hemagglutinin, and relative application. The sequence of the spatial conformation mimic antigen epitope is SEQ ID No. 1. The invention utilizes anti-recombination HA1 rabbit serum IgG selection phage 12 peptide library, uses phage ELISA, DNA sequencing, western bolt, and software analysis to analyze selected colony, and selects the small peptide from random phage 12 peptide library, which can be specially combined with anti-recombination HA1 rabbit serum IgG. The inventive spatial conformation mimic antigen epitope can prepare diagnostic reagent box of H3-type influenza virus.

The invention belongs to the technical field of biomedicine, and particularly relates to the application of a molecular target in the prognosis prediction and treatment of esophageal squamous cell carcinoma. The molecular target is a long-chain non-coded RNA AGPG. The expression level of an AGPG gene in esophageal squamous cell carcinoma tissues is detected through RT-PCR, RNA scope in-situ hybridization, in-situ hybridization, a chip or a high-throughput sequencing platform. The expression level of a target protein PFKFB3 corresponding to the AGPG gene in the esophageal squamous cell carcinoma tissues is detected through immunoblot and an immunohistochemical technology. A biologically active drug is successfully developed, the biologically active drug comprises an inhibitor of the long-chain non-coded RNA AGPG and an inhibitor PFK15 of a PFKFB3 protein, and the biologically active drug can be effectively used for the prognosis prediction and treatment of the esophageal squamous cell carcinoma. A novel molecular target is provided for the prognosis prediction and treatment of the esophageal squamous cell carcinoma by a researching outcome, and a novel treatment strategy is providedfor the clinical treatment of the esophageal squamous cell carcinoma.

The present invention relates to medicine molecular biology and immulogical detection technology, and is one kind of 1N-end polyclonal antibody PAPC1 of anti-human polycystic protein-1. According to cDNA sequence of human polycystic kidney disease-1 gene and the molecular structure of its encoded product, the gene is cloned to cDNA encoding PC-1 and its prokaryotic expression carrier is constituted by means of molecular biology technology. N-end fusion protein of PC-1 is induced in colibacillus and used as antigen for immunizing animal to prepare antiserum and is purified to obtain polyclonal antibody PAPC1. Comprehensive performance test shows that the antibody has stable performance, high sensitivity and powerful specificity in immunohistochemistry, enzyme-linked immunosorbent assay, immunoblotting and immunoprecipiation experiments, so that it may be used in preparing reagent for polycystic kidney disease mechanism research.

The invention discloses an anti-paralichthys olivaceus immunoglobulin D monoclonal antibody. The anti-paralichthys olivaceus immunoglobulin D monoclonal antibody is secreted by hybridoma cells of a hybridoma cell strain JF-IgD, with the collection No.: CCTCCNO: C2014159, the collection unit: China Center for Type Culture Collection, the collection address: Wuhan University in Wuhan, Hubei of China, and the collection date: September 2, 2014. Indirect enzyme-linked immune reaction experiment results show that the anti-paralichthys olivaceus immunoglobulin D monoclonal antibody can be specifically bound with recombinant flounder IgD heavy chain proteins; immunoblotting results show that the anti-paralichthys olivaceus immunoglobulin D monoclonal antibody can specifically recognize recombinant protein encoded by flounder IgD heavy chain genes delta1-delta 4 and flounder natural IgD heavy chain proteins with the molecular weight of 120kDa. The anti-paralichthys olivaceus IgD monoclonal antibody is prepared, and an important tool is provided for detecting flounder IgD protein molecules and IgD surface positive (IgD +) lymphocytes.