40 results about "New zealand white" patented technology

The invention discloses an antibody preparation method through which various organic phosphorus pesticide residues can be detected and the application, which belongs to the technical filed of agriculture. The antibody preparation method adopts the steps that firstly, O,O-diethylthiophosphoryl chloride is used to react with 4-aminobutyric acid to synthesize into general hapten O, O-diethyl-N-(3-carboxy-propyl)-by-sulfur Phosphoramidate DENP which comprises various organic phosphorus pesticide common structures-O, O-diethyl thiosulfate phosphoryl; secondly, the general hapten O, O-diethyl-N-(3-carboxy-propyl)-by-sulfur Phosphoramidate DENP is respectively coupled with bovine serum albumin BSA and chicken ovalbumin OVA by adopting carbodiimide method, to prepare general immunogen DENP-BSA and general coatingen DENP-OVA; and thirdly, multi-clonal antibody is prepared through immunized New Zealand white rabbits, the obtained antibody has obvious specificity to various organic phosphorus pesticides, such as thimet, demeton, disulfoion, omethoate, phoxim and quinalphos and can be used for the immune detection of the organophosphorus pesticides.

The invention discloses a diethoxy thiophosphate organophosphorus pesticide antibody and a preparation method and the application thereof. The antibody of the invention is prepared by introducing appropriate active arms to a diethoxy thiophosphate structure to prepare immunity hapten, coupling the hapten with bovine serum albumin to prepare immunity antigen and immunizing the originally immune New Zealand white rabbits; and a multi-residue ELISA detection method of the diethoxy thiophosphate organophosphorus pesticide is established with coatingen. Based on the detection method established by the antibody, more than 12 types of the diethoxy thiophosphate organophosphorus pesticide can be detected simultaneously, the limit of detection is 0.0002-3.938mug/ml and the detection method is suitable for on-site supervision of pesticide residues and rapid screening with high flux.

The invention discloses preparation and application of a tumor marker calreticulin detection kit, and belongs to the fields of genetic engineering and clinical diagnosis. A monoclonal antibody and a polyclonal antibody of the CRT (calreticulin) are prepared by recombinant CRT genetic immune BALB/c mice and new Zealand white rabbits; and an ELISA (enzyme-linked immunosorbent assay) kit with high specificity and sensitivity of which the lowest CRT content is 0.3125ng/ml and the detection limit is 40-0.3125ng/ml is built. The collected clinical sample is detected through the prepared ELISA kit; the result shows that CRT in blood serum of a patient with lung cancer is obviously higher than that in normal blood serum; significant statistic difference exists (P is smaller than 0.01); and an experimental method is provided for diagnosis of the clinical patients with lung cancer.

The invention relates to a polyclonal antibody of dimethachlon and preparation and application thereof. The polyclonal antibody is prepared by the following steps: coupling hapten and bovine serum albumin BSA to prepare immune antigen; and immunizing New Zealand white rabbits. The polyclonal antibody is characterized in that: the hapten is dimethachlon-2-glutaric acid semi-ester; the dimethachlon-2-glutaric acid semi-ester is coupled with bovine serum albumin by an active ester method to synthesize the immune antigen; and the immune antigen is used for the New Zealand white rabbits so as to obtain the polyclonal antibody. The polyclonal antibody detects the residue of dimethachlon pesticide in a specimen by adopting an indirect competitive ELISA method; and envelope antigen used in the detection is a conjugate of the hapten and ovalbumin by the active ester method. The invention has the advantages that: the preparation method for the polyclonal antibody of the dimethachlon is simple; the polyclonal antibody can be used for quickly detecting the residue of the dimethachlon pesticide in the specimen; and compared with the prior equipmental analysis, the detection method has the advantages of simpleness, quickness, low detection cost, suitability for quickly screening a great number of samples, and the like.

The invention discloses a preparing method and a detecting method for MRJR1 antibody pairs in honey and a kit. The amino acid sequence of the specific polypeptide M4 of MRJR1 is IKEALPHVPIFD, the IKEALPHVPIFD is shown in SEQ ID NO:1; the amino acid sequence of the specific polypeptide M5 of the MRJR1 is SGEYDYKNNYPSDID, and the SGEYDYKNNYPSDID is shown in SEQ ID NO:2; New Zealand white rabbits are immunized. An ELLSA double-antibody sandwich method includes the following steps that a caught antibody SP-1 is coated, sealed and washed, an antigen is added, an enzyme-labeled detecting antibody HRP-SP-2 of an is added, washing is carried out, a color rendering substrate is added, a color rendering stopping solution is added, and the light absorption value of 450 nm-630 nm of a sample is measured through an enzyme-labeling instrument. The antigen is to-be-detected honey or pure honey of a known honey source, and the quality of the to-be-detected honey is judged by comparing the light absorption value of the MRJR1 of the to-be-detected honey and the light absorption value of the MRJR1 of the pure honey of the known honey source. According to the preparing method, the detecting method and the kit, a new reliable rapid detecting method is provided for the quality of the honey and detection of adulteration amount, and the detecting method can be used for honey-product-quality control of honey-product-quality supervision departments and honey-product-quality control of trade processing enterprises.

The invention relates to a methoxy-organophosphorus pesticide broad-spectrum specific polyclonal antibody and the application; the invention is characterized in that the antibody is obtained through the following steps: phosphate is taken as the semi-antigen and coupled with bovine serum albumin BSA to prepare an antigen, and immunise New Zealand white rabbits to prepare the polyclonal antibody. The polyclonal antibody is used to detect the methoxy-organophosphorus pesticide residue through indirect competitive ELISA; the coating antigen used in the detection is a conjugate between hapten and OVA through active ester or mixed anhydride method. The methoxy-organophosphorus pesticide broad-spectrum specific polyclonal antibody has the advantages that the broad-spectrum specific antibody can detect a variety of methoxy-organophosphorus pesticide in vegetables and other foods at the same time so as to significantly save the sample testing time and cost, thereby fully demonstrating the rapid and low-cost characteristics of immunoassay technology.

The invention discloses an ELISA (Enzyme Linked Immunosorbent Assay) test kit for testing difenoconazole residue and a test method. Chloroethane is utilized to substitute hydrogen atoms of three sites on a benzene ring in a difenoconazole molecular structure with ethyl groups, which are oxidized into carboxyl groups, afterwards, the mixed anhydride method is utilized to connect a hapten onto a carrier, so that an artificial antibody is formed, the carrier is bovine serum albumin, an artificial difenoconazole antigen is utilized to immunize New Zealand white rabbits to prepare an antibody, and along with related agents, such as standard difenoconazole, washing concentrate, substrate color-developing solution and stop solution, the antibodies compose the ELISA test kit. The main steps of a test include: sample preprocessing, kit testing, and test result analysis. Compared with a variety of current conventional test methods applied widely, the enzyme linked immunosorbent assay method disclosed by the invention has advantages, such as low cost and high testing sensitivity, and an effective method is provided for difenoconazole residue testing.

The invention provides a brachypodium distachyon functional centromere histone CENH3 and application. Firstly, the nucleotide sequence of a brachypodium distachyon functional centromere histone CENH3 gene is predicted through the bioinformatics sequence comparison method, a polypeptide is synthesized according to a segment of specific sequence of the N end of an amino acid sequence encoded by the nucleotide sequence, New Zealand white rabbits are conventionally immuned through the polypeptide, antiserums are prepared, and a CENH3 protein antibody is obtained through antigen affinity purification. The CENH3 antibody can be used for immunostaining experiments, physical positioning of centromere and chromatin immunoprecipitation (ChIP) experiments, the functional DNA sequence of a brachypodium distachyon functional centromere is obtained, positioning of centromere genome genetic maps and physical maps is carried out, and important tools and information are provided for research of centromere formation and function exercise mechanisms and building of artificial chromosomes.

The invention relates to an immune modulator for treating arteriosclerosis. Aiming at antigen peptides's weak immunogenicity, a series of epitopes peptides of antigens (SEQ ID NO.1) originating from heat shock protein (HSP60) are inserted into the downstream of Cholera toxin B subunit(CTB), and fusion expression of antigens epitopes peptides and CTB is realized. Produced fusion protein (CTB-p277) and Heat shock protein 65 (HSP65) are coupled chemically, without adjuvant, and directly used immunization. The coupled protein can stimulate body to produce high titer antibody aiming at P277 through mucosal immunity pathway, obviously inhibit occurrence of New Zealand white rabbits's arteriosclerosis, and can have a function for treating arteriosclerosis.

The invention belongs to the technical field of medical biology, and relates to a codon-optimized H3HA/XJ3-07 gene and a nucleic acid vaccine thereof. In the invention, according to an HA protein sequence of an equine A-type influenza virus A/Equine/Xinjiang/3/07 (H3N8), an H3HA/XJ3-07 gene segment which can express a corresponding protein is chemically synthesized after codon optimization, and the segment is cloned into a nucleic acid vaccine vector pJW4303 to construct an equine influenza H3HA nucleic acid vaccine; and further, HA genes are modified, an H3-XJ HA natural signal peptide is replaced by a human tPA signal peptide or the gene segments of part of the HA genes are removed to make the HA genes only express an extracellular domain of the HA protein. The experimental result shows that three H3HA nucleic acid vaccines can be expressed in a 293T cell with high efficiency, and can induce the generation of specific anti-H3HA antibodies in New Zealand white rabbits.

A preparation method of a grass carp ATG12 polyclonal antibody comprises the steps: firstly, taking grass carp liver RNA as a template, carrying out reverse transcription to synthesize cDNA, then taking the cDNA as a template, amplifying by PCR with primers and recovering a target fragment, and then constructing an ATG12/pGEX-4T-1 prokaryotic expression vector; transferring the ATG12/pGEX-4T-1 prokaryotic expression vector into competent cells of escherichia coli DH5[alpha], treating, then carrying out ultrasonic crushing and centrifugation to obtain a supernatant and a precipitate, and then purifying the precipitate to obtain purified ATG12 fusion protein; and finally, fully mixing and emulsifying the purified ATG12 fusion protein as an antigen with a Freund's complete adjuvant with the same volume with the ratio of 1:1, immunizing pure New Zealand white rabbits, extracting heart blood, and separating serum, to obtain the ATG12 polyclonal antibody. The antibody has the advantages of high specificity, low cost and short period, and lays the foundation for obtaining the commercially available ATG12 antibody.

The invention relates to a method for in vitro capacitation of rabbit sperms and a nutrient solution formula. In the invention, 10 mum mol/L of EGCG is added in a BO capacitation liquid for sperm in vitro capacitation treatment, and 10 mum mol/L of EGCG is added in the BO capacitation liquid, the formula of the a BO capacitation liquid comprises the following components by weight: 6.5453mg/mL of NaCl, 0.2997mg/mL of KCL, 0.1145mg/mL of NaH2PO4.H2O, 0.1057mg/mL of MgC12.6H2O, 0.330mg/mL of CaCl2.2H2O, 2.518mg/mL of glucose, 4% of 0.2% phenol red, 100IU/mL of penicillin, 100IU/mL of streptomycin, 3.1080mg/mL of NaHCO3, 0.1380mg/mL of sodium pyruvate, 20mum g/mL of heparin and 20mg/mL of BSA. The method for in vitro capacitation of rabbit sperms overcomes the defect of mass oxyradical accumulation generated by sperm metabolism due to large sperm density during a current sperm in vitro capacitation process. The method uses adult New Zealand white male rabbit, different concentration EGCG (0mumM, 10mumM, 15mumM, 20mumM and 25mumM) can be added in the BO capacitation liquid, influence of the BO capacitation liquid to the rabbit sperm capacitation effect is researched so that optimum EGCG addition concentration is obtained, and thereby a rabbit in vitro capacitation culture system with high efficiency is established.

The invention discloses a wheat yellow striate virus N gene recombination expression protein, a method for preparing polyclonal antibodies and application thereof. The method for preparing the polyclonal antibodies includes steps of synthesizing genes by optimization codons according to sequences of N genes of WYSV (wheat yellow striate viruses) for the first time and subcloning the genes into target vectors pET-30a for expressing Escherichia coli; carrying out prokaryotic expression on recombinant proteins and immunizing New Zealand white rabbits to obtain the WYSV-N protein polyclonal antibodies. The wheat yellow striate virus N gene recombination expression protein, the method and the application have the advantages that as shown by Western blot detection, the antibodies prepared by theaid of the method can be specifically bound with toxic insect samples, and accordingly the high specificity of the obtained antibodies is illustrated; whether vector insects carry the WYSV or not canbe quickly detected by the aid of immunofluorescence detection processes under laboratory conditions, and accordingly a foundation can be laid for research on WYSV prediction and forecast and psammotettix striatus L. and WYSV interaction mechanisms.

The invention relates to the field of biology and aims to provide application of high mobility group box (HMGB) protein and antibody to the preparation of an ostrea rivularis anti-infection immune preparation. The amino acid sequence of the protein is shown as the SEQ ID NO:2; the coding frame nucleotide sequence of the gene is shown as the SEQ ID NO:1; the application of the protein to the preparation of the ostrea rivularis anti-infection immune preparation is to immunize New Zealand white rabbits by using the protein so as to obtain polyclonal antiserum. According to the invention, the ostrea rivularis HMGB gene coding frame full sequence is obtained; the soluble HMGB protein product is expressed and purified by establishing a prokaryotic expression vector; and the polyclonal rabbit antiserum is prepared. The polyclonal rabbit antiserum has the effect of inhibiting inflammatory response caused by ostrea rivularis pathogeny RLO and gram negative bacteria LPS as well as necrobiosis and apoptosis.The invention relates to the field of biology and aims to provide application of high mobility group box (HMGB) protein and antibody to the preparation of an ostrea rivularis anti-infection immune preparation. The amino acid sequence of the protein is shown as the SEQ ID NO:2; the coding frame nucleotide sequence of the gene is shown as the SEQ ID NO:1; the application of the protein to the preparation of the ostrea rivularis anti-infection immune preparation is to immunize New Zealand white rabbits by using the protein so as to obtain polyclonal antiserum. According to the invention, the ostrea rivularis HMGB gene coding frame full sequence is obtained; the soluble HMGB protein product is expressed and purified by establishing a prokaryotic expression vector; and the polyclonal rabbit antiserum is prepared. The polyclonal rabbit antiserum has the effect of inhibiting inflammatory response caused by ostrea rivularis pathogeny RLO and gram negative bacteria LPS as well as necrobiosis and apoptosis.

The invention relates to a rapid chemiluminiscence detection kit for quinolone drugs and a testing method of the kit. The kit is internally provided with a luminescent plate, a quinolones polyclonal antibody treated by a chemical primer, a goat-anti-rabbit antibody marked by horseradish peroxidase, a quinolones-series standard solution, a concentrated phosphate buffer, a concentrated washing solution and a luminescent solution, wherein the luminescent plate is provided with a coating antigen; the coating antigen is obtained by coupling a quinolone drugs dry hapten and OVA by adopting a mixed anhydride method; the quinolones polyclonal antibody treated by the chemical primer is obtained by taking a conjugate as an immunogen to immunize New Zealand white rabbits and the conjugate is prepared by coupling norfloxacin, an ofloxacin derivative and the OVA; the lowest detection limit of a chemiluminiscence enzyme-linked immunosorbent assay detection kit can be up to 0.1ng/ml, a linear detection range is 0.1ng/ml-8.1ng/ml, the in-plate variable coefficient is less than 20% and the recovery rate in a water sample ranges from 78% to 118%.

The invention provides a preparation method of rabbit anti ovotrasferrin allergen polyclone antibody and an immunoassay method thereof. The preparation method comprises the steps that ovotransferrin rabbits are directly immune to New Zealand white rabbits, after immunization is conducted for five times, whole blood is taken from femoral arteries, and ovotransferrin antibody serum is obtained; the antibody peridium amount is optimized, the dilution ratio of the antibody, confining liquid and the PH value of a sample buffer solution are detected, and an indirect competitive ELISA method among the ovotransferrin is established. The antibody prepared through the method is high in potency and good in inhibition rate, and the established method is good in sensitivity and accuracy.

The invention relates to a method for repairing the rabbit achilles tendon injury through continuous stretching stress stimulation. The method comprises the steps that a male New Zealand white rabbitis adopted, and through hind limb achilles tendon dividing operation, a rabbit achilles tendon acute breakage animal model is established; at functional positions, ankle joints of the rabbit are fixedthrough gypsum to limit moving of the ankle joints of the rabbit, continuous stress stimulation is given to the achilles tendon breakage section, and the animal model for achilles tendon repairing under early-stage continuous stress stimulation is constructed; through general observation and microscopic observation, the early-stage histological change after rabbit achilles tendon acute breakage repairing operation under fixing of the functional positions of the ankle joints is defined; through the ELISA method, protein expression situations of I-type collagen, TGF-beta1, bFGF, VEGF, PDGF andIGF-1 in repair tissues are detected; and by detecting the maximum strength, the maximum load and the rigidity index, the biomechanical characteristics of the repair tissues after early-stage continuous stress stimulation is given in the rabbit achilles tendon acute breakage postoperation repairing process are evaluated.

The invention relates to the technical field of biology, in particular to expression of a human hepatitis B vaccine (namely hepatitis B virus (Hepatitis B virus, HBV) surface antigen protein) throughplants. The plants such as lettuces are used as an effective expression platform for producing recombinant protein, and the human Hepatitis B vaccine is expressed by a simple and effective agrobacterium tumefaciens mediated vacuum osmotic method. The expression system determines that plant foreign protein can be collected after agrobacterium tumefaciens is subjected to infection for 4d. By an SDS-PAGE method, successful expression of the recombinant human Hepatitis B vaccine protein is determined. After expressed recombinant protein is prepared into virus like particles through self-assembly,New Zealand white rabbits can be immunized, and obtained serum can prove that the human Hepatitis B vaccine expressed by the plants has biology activity by an enzyme-linked immunoadsorbent assay method (ELISA) and a pseudovirion granule neutralization test.

A harmful red tide organism cell surface specific antibody and a preparation method thereof relate to an antibody. The invention provides a harmful red tide organism cell surface specific antibody and a preparation method thereof. The antibody is an immunoglobulin which is produced by the plasma cells differentiated from B cell and can be combined and reacted with corresponding antigen in a specific way under the incentive of a red tide algae surface antigen substance. Firstly, East China Sea chain prorocentrum Alexandrium whole-cell surface antigen which is not contaminated by the protein in the cell is prepared and then mixed with complete Freund-type adjuvant, later, the mixture is injected into New Zealand white rabbits, and then antigen is injected and injection is enhanced; New Zealand white rabbit ear vein blood is collected 3 to 8 days before the serum antibodies are collected, and later the generation condition of antibodies is detected; the whole rabbit blood is collected 3 to 5 days after the last injection; and after standing and centrifugation, the serum is collected and then packaged and stored. Aiming at the red tide of algae type, corresponding red-tide algae species can be rapidly detected by using the antibody; and the detection limit can reach more than a dozen cells.

The invention relates to a malachite green oxalate artificial antigen, in particular to a method for preparing a malachite green oxalate artificial antigen. The method sequentially includes steps of synthesizing prepared artificial hapten and hapten 4-MGC (methyl glycol chitosan); and synthesizing a prepared malachite green oxalate artificial antigen, malachite green oxalate and BSA (bovine serum albumin). The method for preparing the malachite green oxalate artificial antigen has the advantages that a synthesizing process is advanced, the specificity is high, as shown by detection results obtained after the prepared malachite green oxalate artificial antigen is used for immune New Zealand white rabbits, the titer of immune sera of the malachite green oxalate artificial antigen is 1:72000, the malachite green oxalate artificial antigen is completely applicable to immunoassay, and a convenient, speedy and accurate channel for detecting malachite green oxalate is provided.

The invention discloses a preparation method and application of a forchlorfenuron artificial antigen and polyclonal antibody. The preparation method mainly includes the steps of firstly, using Friedel-Crafts reaction to synthesize artificial forchlorfenuron hapten; secondly, using the artificial forchlorfenuron hapten as the raw material, and using a carbodiimide method to couple the artificial forchlorfenuron hapten with bovine serum albumin to obtain the forchlorfenuron artificial antigen; thirdly using the synthesized artificial antigen to immunize New Zealand white rabbits, taking blood, centrifuging to obtain antiserum, namely the anti-forchlorfenuron polyclonal antibody. The preparation method and the application have the advantages that the problems that a traditional physicochemical analysis method is complex, high in cost, slow in analysis and the like are solved, and the immunoassay technology which is simple, fast and sensitive is provided; the prepared antibody is good in specificity and sensitivity, simple in synthesizing method, applicable to the fast immune detection of forchlorfenuron and good in application value.

The invention relates to a method for preparing an enzyme-labeled antibody for tilapia IgM detection, which belongs to the field of biotechnology. Tilapia serum is collected, and protein G is used for affinity purification to obtain tilapia IgM antibody, and then the tilapia IgM New Zealand white rabbits were immunized with antibodies to obtain rabbit anti-tilapia IgG antibodies, and the rabbit anti-tilapia IgG antibodies were labeled with horseradish peroxidase to obtain enzyme-labeled secondary antibodies. Compared with the prior art, the present invention has beneficial effects: the present invention adopts the method of affinity chromatography to prepare high-concentration rabbit anti-tilapia IgM polyclonal antibody, and carries out horseradish peroxidase labeling and labeling after purification Determination of antibody concentration, the titer is greater than 1:128000, and it is used in the immune reaction through appropriate dilution to detect tilapia IgM, which is helpful for the large-scale prevention and control of tilapia streptococcosis.