63 results about "HIV Western Blot" patented technology

The invention provides a chemiluminiscence diagnostic kit for sensitization allergens and a preparation method thereof. The diagnostic kit comprises a microporous plate coated with the sensitization allergens, a horseradish peroxidase-labeled anti-human IgE (immunoglobulin e) antibody, chemiluminiscence substrate solutions A and B, and a washing solution, wherein the sensitization allergens comprise one or more of an insect allergen, a pollen allergen, a fungus allergen and a food allergen. According to the diagnostic kit, the difference of the sensitivities of different proteins in one diagnostic kit is overcome, and the detection sensitivity of each allergen is enhanced to the maximum extent by repeatedly testing and adjusting the coating amount of the allergens, the concentration of the enzyme-labeled antibody and the formulae and the concentrations of the chemiluminiscence substrate solutions. The chemiluminiscence diagnostic kit is higher in detection sensitivity, safe, reliable, simple and convenient to operate and low in cost in comparison with the ELISA (enzyme linked immunosorbent assay), western-blot and RAST (radioallergo-sorbent test) diagnostic kits for the allergens.

The invention provides a monoclonal antibody TBCD6 resistant to mycobacterium tuberculosis culture filter protein (CFP) 10. A preparation method of the monoclonal antibody is characterized by immunizing the mice with the specific antigenic peptide, collecting sensitized B lymphocyte of the mice to undergo fusion with the myeloma cells of the mice, judging positive clones by utilizing enzyme-linked immunosorbent assay (ELISA) and western blotting, building a hybridoma cell system TBCD6 which secretes the monoclonal antibody, amplifying, culturing and injecting the positive cells to the peritoneal cavities of the mice of the same strain, using the mice to induce ascites containing the antibody and obtaining the monoclonal antibody through collection and affinity purification of the antibody. The invention provides application of the monoclonal antibody to detection and quantitative analysis of the mycobacterium tuberculosis CFP 10.

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 discloses recombinant neutrophil gelatinase associated lipocalin and a preparation method thereof. The preparation method of the recombinant neutrophil gelatinase associated lipocalin antibody belonging to the technical fields of biological science and transthyretin comprises the following steps: analyzing an NGAL (neutrophil gelatinase associated lipocalin) epitope; synthesizing the recombinant NGAL; separately purifying the recombinant NGAL; preparing an anti-rabbit recombinant NGAL antibody; collecting and separating to obtain an antiserum purified antibody containing the antibody, so as to obtain the anti-NGAL antibody. By adopting the preparation method, the problem of low yield in preparation of a multi-antibody by a natural full-length protein is solved; meanwhile, the antibody prepared by the preparation technology disclosed by the invention has good specificity; the problem that the traditional neutrophil gelatinase associated lipocalin antibody cannot completely meet a western blot immunohistochemical experiment and an enzyme-linked immunosorbent assay is also solved.

The invention provides a recombination lactic acid galactococcus method of expessing pig transmissible gastroenteristis virus mutation S protein, which comprises the following steps: designing double primers according to the total gene order of pig Transmissible gastroenteristis virus S protein and gene fusion characteristic of expressing the carrier plasmid; proceeding PCR; getting the 2007bp destination fragments of four main antigen sites with TGEV S gene; connecting with the carrier plasmid PNZ8112 of secretory expression; getting into the host lactic acid galactococcus NZ9000 cell by electric conversion; expressing under the inducing of nisin. The invention constructs the expression carrier system of coronaviridae TGEV S mutein lactic acid galactococcus, which expresses about 66KDa destination protein of four main antigen sites with TGEV. Immunological ink mist experiment and indirect immunological fluorescein test indicate that the exogenesis protein is able to react with TGEV immunological serum and the recombination S protein has the same antigenic as TGEV natural antigenic.

The invention discloses a preparation method of HPV16aE7/huhsp70 fusion protein. According to the preparation method, pRb binding sites Cys24Gly and Glu26Gly, CK II sites Ser31Gly and Ser32Gly as well as zinc finger binding domains Cys58Gly and Cys91Gly of wild type E7(wE7) protein are mutated, meanwhile, CTL epitope 11-20aa and 49-57aa as well as Th accessory cell epitope 30-67aa with E7 specificity are added at protein N end and C end of the protein E7 respectively, transformed and optimized HPV16 mE7 gene is obtained, a pET-30a-mE7 prokaryotic expression vector is built, a pET-30a-huhsp70 prokaryotic expression vector of huhsp70 is built, mE7 and huhsp70 are fused, the prokaryotic expression vectors are inserted, prokaryotic expression plasmid pET30a-mE7/huhsp70 is built, protein induction expression is performed in escherichia coli BL21(DE3), expression activities of three kinds of protein are identified with SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) and western blotting, and then soluble analysis and purification of recombinant protein are performed. The preparation method has the advantages of simplicity, feasibility, high universality and good tumor treatment effect.

The invention discloses a liquid-chip HIV-antigen antibody combined detection kit and a detecting method and belongs to the field of immunoassay. The kit comprises microspheres respectively covered with HIV gp41, p17, p24, p31, p66 and gp120 antigens, a biotinylated anti-human secondary antibody, an SA-PE solution, microspheres covered with a p24 monoclonal antibody, and a monoclonal antibody labeled with fluorescein and matching the p24 monoclonal antibody. Combined detection for six HIV antibodies and a P24 antigen can be performed simultaneously through a small amount of a sample. Compared with an immunoblotting method, the kit and the detecting method are high in sensitivity and specificity. Through the kit and the detecting method, the HIV antibodies can be detected timely in the earlier period of infection, thus achieving early detection and early intervention of HIV infection, reducing HIV transmission and ensuring safety of blood transfusion. The kit and the detecting method have a wide application prospect.

The invention provides a monoclonal antibody TBEF3 of tubercle bacillus-resistant secretory antigen target protein in early stage. The monoclonal antibody TBEF3 is prepared by the following steps: selecting a specific antigen peptide mouse; collecting sensitized mouse B lymphocyte and mouse myeloma cell to fuse; judging positive clones by using an enzyme linked immunosorbent assayelisa and an immunoblotting testing method; establishing a hybridoma cell line TBEF3 for secreting monoclonal antibody of tubercle bacillus-resistant secretory ESAT-6 in early stage; carrying out multiplication culturing on the positive cells and injecting to congenic mouse enterocoelia for induction of generation of ascitic fluid containing antibody; and carrying out collection and antibody affinity purification to obtain the monoclonal antibody TBEF3 of tubercle bacillus-resistant secretory antigen target protein in early stage. The invention also provides an application of the monoclonal antibody in testing the secretory antigen target protein of mycobacterium tuberculosis in early stage.

The invention discloses a method for detecting specificity of Epstein-Barr viruses (EBV) through an Epstein-Barr virus p18-p23 fused capsid antigen. The method is characterized in that an overlap extension polymerase chain reaction (PCR) technology is adopted; a coding gene p23 and a coding gene p18 are fused in vitro by a coding sequence of an intermediate head (Gly4Ser)3; a fused protein is expressed and the expressed fused protein is purified and is subjected to specificity identification through an immunoblotting technology; the purified fused protein is coated on an enzyme-linked immuno sorbent assay (ELISA) reaction plate and component content and reaction conditions are optimized; commercial horseradish peroxidase-labelled goat anti-human IgM and IgG and a series of other reagents are prepared; and EBV-viral cuspid antigen (VCA) specific IgM and IgG indirect ELISA diagnostic kit is obtained by assembling.

The invention discloses a polypeptide for inhibiting proliferation of tumor cells, which is of a fragment with maximum length of 145 amino acid residues at 46-100 position from N-terminal, comprising ribosomal protein RPL26. The reporter gene experiments, Western blotting, co-immunoprecipitation and cell proliferation detection experiments confirm that RPL26 as well as MDM2 and p53 form a complex, and the combination of RPL26 and MDM2 can obviously reduce the ubiquitin ligase activity of MDM2, thereby improving the stability and transcriptional activation of p53. The overexpression of RPL26 can obviously inhibit the proliferation of the tumor cells, and the function is related to the activity of p53. RPL26 can be assisted for the clinical treatment of cancer and has great significance.

The invention provides a tumor marker AMP-18 antigen polypeptide which has an amino acid sequence shown as SEQ ID No.1 in a sequence table. The invention also discloses a method for cloning and expressing the protein by a gene and carrying out gastric cancer biological behavior judgment and an animal test, wherein two different test technologies of immunoblot and confocal are applied to obtain the same conclusion that the AMP-18 can be combined with a gastric mucosa epithelial cell and is the autocrine protein of the cell. Because the AMP-18 has favorable tumor inhibiting effect, the AMP-18 antigen polypeptide can be used for the application in preparing a medicament for treating a tumor.

The invention provides a monoclonal antibody pSer86 of Tip60, as well as the preparation method and the application of the pSer86 monoclonal antibody, and relates to a novel cell strain and the monoclonal antibody. The cell strain is a monoclonal antibody cell strain pSer86-13MAb obtained by phosphorylating the No. 86 serine of Tip60; the monoclonal antibody obtained by phosphorylating the No. 86serine of Tip60 is an antibody generated by the monoclonal antibody cell strain pSer86-13MAb obtained by phosphorylating the No. 86 serine of Tip60; the antibody excreted by the cell strain can be used for detecting ELISA (enzyme linked immunosorbent assay), immunoblotting, co-immunoprecipitation, immunofluorescent staining and the like; and the antibody can identify not only exogenous overexpression phosphorylated Tip60, but also cell endogenous phosphorylated Tip60, provides convenience for developing and researching phosphorylated Tip60, and provides the first monoclonal antibody with Tip60 of which the No. 86 serine is phosphorylated.

The invention provides an anti-FtsH2 protein monoclonal antibody and an application thereof. The antibody is secreted from a mouse hybridoma cell line 30749-22 and can be used for preparing a kit for screening a plant material with a stress resistance character. According to the anti-FtsH2 protein monoclonal antibody, a recombinant FtsH2 protein and a natural FtsH2 protein expressed in wheat leaves can be specifically combined and identified. The antibody is an IgG1 antibody, and has extremely high specificity and sensitivity when combined with FtsH2 protein; the monoclonal antibody is capable of carrying out qualitative and semi-quantitative analysis on FtsH2 protein in plant leaves through a Western blotting method, and is used for identifying genetic modified plants, thereby screening a plant material with high stress resistance.

The invention discloses a preparation method of an SETD3 antibody for immunofluorescence assay. The method comprises the following steps: immunizing rabbit with protein expressed through full-length SETD3 which is labelled with N-terminal 6xHis, and extracting serum of the immunized rabbit; and purifying the serum of the rabbit with the full-length SETD3 protein which is labelled through GST that is coupled with Affi-Gel. The SETD3 antibody is applicable to western blot (WB), immunofluorescence (IF), immunohistochemistry (IHC) and immunoprecipitation (IP), and is high in titer and specificity.

Compositions, formulae, devices and methods for the detection of target microorganisms, such as by visual immunoprecipitate assay, enzyme linked immunoassay, chemiluminescence, immunoblotting, or similar detection technology, wherein detection requires the discrimination among closely related genera, species and strains of antigenically related microorganisms based on immunological reactivity of a highly conserved antigen epitopes with a reagent system comprised of an antibody linked to a detecting reagent. The invention permits a detectable event to occur by exposing inaccessible but highly conserved and specific antigen epitopes to the detecting reagent. Exposure of such antigen epitopes without inactivating microbial metabolism allows for specific detection.

The invention provides a western blot kit for specific IgG antibodies of syphilis and a preparation method thereof, relating to a kit. The kit is provided with a carrier plate, a cellulose nitrate membrane, a detection line of the specific IgG antibodies of the syphilis and a control line, wherein the detection line and the control line are arranged on the cellulose nitrate membrane in sequence; specific recombinant antigens of the syphilis are coated at the detection line of the specific IgG antibodies of the syphilis, and human IgG antibodies are coated at the control line; and horse radish peroxidase is marked on anti-human gamma-chain antibodies. The preparation method comprises the following steps: preparing the specific recombinant antigens of the syphilis, carrying out sample application on the cellulose nitrate membrane, and then preparing gamma-chain monoclonal antibodies of anti-human IgG specific segments; and marking the horse radish peroxidase on the anti-human IgG specific segments gamma-chain monoclonal antibodies, and then preparing the western blot kit. The invention can be used for detection of the specific IgG antibodies of the syphilis in specimens such as whole blood, blood serum, blood plasma, cerebrospinal fluid and the like. In the invention, the amount of the needed specimens is very small, special instruments are not needed, the detection is simple, convenient and fast, the specificity is strong, the sensitivity is high, the accuracy and the reliability are achieved, and the cost is low.

The invention relates to an insulin-like growth factor (IGF) family protein combined detection chip. The chip includes: glass slides coated by active amino propyl silane; multiple micro-arrays fixed on each glass slide by a chip dot matrix; and gaskets for separating the micro-arrays into different hybrid areas, wherein each of the micro-arrays contains a specific antibody corresponding to an IGF family protein. The chip also includes the following reaction reagents: an biotin-marked detection antibody mixed solution, a closed solution which adopts a phosphate buffer solution containing 5% bovine serum albumin, a standard protein mixture, and streptavidin marked by adopting a fluorescent dye HyLight555 with the excitation wavelength of 550 nm and the emission wavelength of 566 nm. The IGF family protein combined detection chip can complete multi-detection indicator, high-throughput and high-sensitivity detection which cannot be completed by traditional methods such as ELISA, western blot and the like, and is quite suitable for multi-link simultaneous detection of mass target proteins of an IGF-1 signal pathway.

The invention discloses an immunoblotting kit for sjogren syndrome specific autoantibody detection. The kit includes: a blotting membrane strip placed in a reaction tank, an enzyme conjugate, a chromogenic reagent A, a chromogenic reagent B, a stop solution, a dilution solution, concentrated washing liquid, and a standard zone tape. The membrane strip is placed into the reaction tank to react with to-be-tested serum, if the to-be-tested serum contains a corresponding autoantibody, the autoantibody can combine with a corresponding antigen, then the enzyme conjugate and the chromogenic reagents are added, contrast with the standard zone tape is carried out to judge whether the serum contains the sjogren syndrome specific autoantibody: anti-SSA antibody (Ro), SSB (La), salivary gland protein 1 (SP1), carbonic anhydrase 6 (CA6), and parotid secretory protein (PSP). The existence of sjogren syndrome specific autoantibodies and the antibody species and quantity are of important clinical significance for sjogren syndrome diagnosis and differential diagnosis.

The invention discloses a monoclonal antibody of anti-human BAP31 protein, a preparation method and the application thereof, belonging to the preparation of antibody-containing medical preparation or the monoclonal antibody. The monoclonal antibody of the anti-human BAP31 protein is generated by a hybridoma cell with the collection number of CGMCC No. 3271. The breast adenocarcinoma MCF-7 immuno-BALB/c mouse is adopted, spleen and myeloma cell of the mouse are taken to make cell fusion, the MCF-7 cell is utilized for screening, and then a monoclonal antibody library is formed by the obtained positive clones; and the antigen of the monoclonal antibody derived from the antibody library can be identified to be BAP31 by immuno-precipitation and a mass spectrum method. A variable region sequence for identifying the antibody can be determined, and the antibody can distinguish the BAP31 protein in a specific way and has specific binding capacity with the BAP31 protein. The invention is applicable to the detection of immunohistochemistry, immuno-fluorescence, immuno-blot and immuno-precipitation of the molecule.

The invention discloses an inflammatory myopathy SAE1 self antibody non-radioactive detection method and an application thereof. The method comprises the following steps: an inflammatory myopathy SAE1 self antibody non-radioactive mark immunoprecipitation detection method and an immunoblotting detection method which are used for detecting an anti-SAE1 self antibody of the patients with idiopathic inflammatory myopathy; the non-radioactive mark immunoprecipitation detection method comprises the following steps: using Protein G magnetic bead for respectively combining with the SAE1 antibody positive inflammatory myopathy serum or health contrast serum, then performing incubation with over-expressed SAE1 HEK293 cell lysis buffer for antigen adsorption, after elution, performing SDS-PAG gel electrophoresis, and using a Flag antibody for detection; the immunoblotting detection method comprises the following steps: performing SDS-PAG gel electrophoresis on the over-expressed SAE1 HEK293 cell lysis buffer, using SAE1 antibody positive serum a primary antibody, and using rabbit anti human IgG H&l as a secondary antibody for detection. According to the invention, problems of an ELISA kit for detecting SAE1 antibody and immunization immunization blotting membrane without commercialization in our country can be solved, and the problems of high cost of the ELISA kit for self-coating of the SAE1 antigen, high false positive rate and security of a radioactive mark immunoprecipitation method can be overcome.

The invention discloses an anti-TOPK antibody with 74th tyrosine residue phosphorylated as well as a preparation method and an application of anti-TOPK antibody and belongs to the field of biology and disease diagnosis. The antibody is obtained as follows: polypeptides with the amino acid sequence shown in SEQ ID NO: 1 are coupled with carrier protein, a product serves as an antigen, the antigen is used for immunize an animal, polyclonal antibody serums are obtained and purified, and the antibody is obtained. The antibody can be applied to immunoblotting detection of the tumor cell level. Currently, an antibody for detecting Y74 phosphorylation of TOPK is not available in the market. The antibody lays a solid foundation for research of tumor occurrence as well as early diagnosis and prognosis predicted treatment of the tumor.

The invention provides an immunoblotting reagent kit of syphilis specific total antibodies and preparing method thereof, relating to a reagent kit. The reagent kit is provided with a carrier plate, a nitrocellulose membrane, a syphilis specific total antibody testing line and a control line, wherein the syphilis specific total antibody testing line and the control line are orderly arranged on the nitrocellulose membrane; the syphilis specific total antibody testing line is coated with syphilis specific recombinant antigens; the control line is coated with a human total antibody; and horseradish peroxidase is marked with an anti-human Ig antibody. The method comprises the following steps of: firstly preparing the syphilis specific recombinant antigens; applying samples of the nitrocellulose membrane; preparing the anti-human Ig monoclonal antibody; marking the anti-human Ig monoclonal antibody on the horseradish peroxidase; and preparing the immunoblotting reagent kit. The method can be used for testing the syphilis specific total antibodies in the specimens of whole blood, serum, blood plasma, cerebrospinal fluid and the like. The invention has the following advantages: during testing, the needed specimen quantity is small, special instruments are not needed, testing is simple, convenient and rapid; the specificity is strong, the sensitivity is high, testing is accurate and reliable, the cost is low and the invention is widely applied.