53 results about "Sheep serum" patented technology

The invention provides a screening culture method for sheep oocytes in vitro, which comprises the following steps: step 1: ovaries which were killed less than half an hour ago are put into saline water at the temperature of 30DEG C to 40DEG C and containing penicillin and streptomycin, washed for 3 to 4 times within 3 hours, and oocytes are picked out; 25 to 30mu mol / L brilliant cresyl blue is put into 35DEG C to 39DEG C water bath to be dyed for 85 to 92min, the cytoplasm is blue and is washed for 3 to 4 times by in vitro maturation fluid, is put into 55 to 78mul / drip in vitro maturation culture fluid by 25 to 30m / drip, and is cultured in 5 percent CO2 by 95 percent; step 2: cumulus cells are removed from oocytes which are maturated in vitro for 25 to 28 hours; IVF washing liquid is used to wash for 2 to 4 times, and is dripped into 50 to 70mu l fertilization fluid by 25 to 30m / drop; semen is unfrozen, the fertilization fluid is moved in, supernatant fluid is centrifugurated for 4 to 5min and removed, the precipitated sperms are added into fertilization fluid drips by the density of 2 to 4*106 / ml and incubated; eggs which are fertilized for 12 to 18 hours are treated in the above step and then moved into a four-hole culture plate to be cultured; and step 3: reagent egg absorption liquid, brilliant cresyl blue maturation liquid, dyeing liquor SOF, the fertilization fluid, the culture fluid and sheep serum are prepared.

The invention discloses an indirect ELISA detection kit for B-type clostridium novyi. The indirect ELISA detection kit takes B-type clostridium novyi bacterium suspension as an envelope antigen, wherein the B-type clostridium novyi bacterium suspension is prepared by the following methods of inoculating B-type clostridium novyi into a culture medium to carry out bacterium-enrichment culture; centrifuging the cultured B-type clostridium novyi culture; and collecting bacterial cells, washing and resuspending the bacterial cells by using a carbonate buffer solution to prepare the B-type clostridium novyi bacterium suspension. The kit disclosed by the invention takes complete clostridium novyi bacterial cells as the envelope antigen, so that whether sheep serum contains an antibody to clostridium novyi or not can be detected at high sensitivity and high specificity and whether sheep are infected with a black disease or not can be judged accordingly. The indirect ELISA detection kit has the advantages that the detection result is high in specificity and good in repeatability, and a diagnosis can be quickly made and emergency treatment measures can be taken after the sheep catch the disease.

The invention discloses a quality control product which comprises a quality control product 1 and a quality control product 2, wherein the quality control product 1 comprises the following components:2-6 parts by weight of factor-rich goat plasma, 2-5 parts by weight of sheep serum and 1-4 parts by weight of fibrinogen mother liquor; the quality control product 2 comprises the following components: 1 to 4 parts by weight of factor-deficient sheep plasma, 5 to 10 parts by weight of sheep serum and 0. 3 to 2 parts by weight of the fibrinogen mother liquor, the fibrinogen mother liquor is prepared by dissolving fibrinogen in the sheep serum, wherein the mass percentage of the fibrinogen is 10%; clotting factor extraction is not needed in the preparation process of the thromboelastography instrument quality control product, preparation steps are simplified, operation is easy, cost is low, results are accurate, and stability is good; test results of the thromboelastography instrument quality control product basically meet that of a quality control product produced by already listed US Haemoscope company, the thromboelastography instrument quality control product can replace the qualitycontrol product produced by the already listed US Haemoscope company, and customer use cost is reduced.

The invention provides an immunofluorescence kit for detecting CEA gene mutation of peripheral blood circulating tumor cells of a non-small cell lung cancer patient and a detection method, and belongsto the technical field of molecular biology. The kit comprises 45mL of a diluent, 1mL of a destaining solution, 0.5mL of a staining solution A, 1mL of a staining solution B, 200[mu]l of methanol, 200[mu]l of 2% PFA, 100[mu]l of 10% goat serum, 100[mu]l of a primary antibody working solution, 100[mu]l of a secondary antibody working solution and a DAPI mounting medium. According to the detection method provided by the invention, the CEA expression condition of a patient suffering from advanced or recurrent non-small cell lung cancer can be detected without obtaining a tissue specimen through needle biopsy. The technology belongs to a minimally invasive technology and can realize real-time detection. According to the method provided by the invention, a false positive result caused by an edge effect possibly generated in the dyeing process can be avoided, the stability is good, the cell loss is reduced, and the detection accuracy is improved.

The invention provides an extraction method for sheep serum albumin. The method comprises the following steps: (1) fresh sheep blood is taken, an anticoagulant accounting for 12% of the original serumvolume is added, after qualified inspection and quarantine, centrifugation is performed at 4 DEG C and 2500 rpm for 15 min, a supernatant is taken and frozen at subzero 20 DEG C, frozen serum is taken, thaws sufficiently at room temperature and is centrifuged at 10000 rpm for 12 min, and a supernatant is taken and is the sheep serum. The extraction method for the sheep serum albumin is short in flow, simple to operate and high in production efficiency, no organic solvent is used in the method, protein aggregation or denaturation is avoided, meanwhile, environmental pollution and unsafe factors in the production process can be reduced, product purity is improved, and the method can be widely applied to the fields of medical treatment, biochemistry and the like.

The invention discloses an antigen for detecting a sheep echinococcosis antibody and a preparation method of an agar diffusion plate, and belongs to the technical field of biological detection. The preparation method of the antigen comprises the following steps: performing enzyme digestion connection on a sheep echinococcosis EG95 protein nucleic acid sequence, and constructing the sheep echinococcosis EG95 protein nucleic acid sequence on a plasmid vector; then performing converting into escherichia coli, constructing an expression strain, performing IPTG induced expression, and performing crushing to obtain an inclusion body; and performing washing, cracking, centrifuging, purifying, dialyzing, concentrating, freeze-drying, and diluting with PBS. The preparation method of the agar diffusion plate comprises the following steps: adding water and PBS into agarose, performing heating, adding polyethylene glycol 6000, MES and sodium chloride, performing cooling, performing punching, and sealing the bottom. The antigen and the agar diffusion plate provided by the invention have the advantages of high sensitivity, strong specificity and strong universality, have good biosafety, are easyfor large-scale production, are especially suitable for clinical detection of sheep serum by grassroots veterinarians, and have wide popularization and application value.

The invention relates to a veterinary drug, and in particular relates to a preparation process of a perfusion agent for treating dairy cow mastitis. The process comprises the steps of extracting active ingredients of scutellaria baicalensis, flos lonicerae and leonurus through methods of water extraction, concentration, acid eduction and the like. The traditional Chinese medicine perfusion agent has no toxicity, irritation or anaphylaxis; the perfusion agent has better treatment effect on clinical mastitis models of milk goats and the best perfusion dose is 10ml per mammary area; the perfusion agent is capable of remarkably enhancing the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in ill goat serum and reducing the contents of nitric oxide (NO) and malonaldehyde (MDA).

The invention provides a method for producing transgenic sheep by injecting lentivirus into perivitelline space; lentiviral vector and virus packaging plasmid are cotransfected into 293T cells and then centrifugated after 48 hours to prepare the concrete of the lentiviral; the concrete of the lentiviral is injected into oocyte perivitelline space which is 24 to 25 hours mature, is integrated with sperms to implement in vitro fertilization, is put into culture fluid after 12 hours to be cultured, and the cleavage rate and the transgenic rate are calculated after 48 hours. Step 1: an oocyte is washed by in vitro maturation fluid for 3 to 4 times, and the 75 to 78um l / drop in vitro maturation fluid which is dripped by 25 to 30m / drop is cultured in 5 percent CO2 by 95 percent; step 2: a cumulus cell is removed from the in vitro matured oocytes, the lentiviral is injected into the cell perivitelline space and is washed by IVF washing liquid for 2 to 4 times, and fertilization fluid is dripped by 25 to 30m / drop; semen is unfrozen, the fertilization fluid is moved in, supernatant fluid is centrifugurated and removed, the sperms are precipitated and added into fertilization fluid drips and incubated; eggs which are fertilized for 12 to 18 hours are treated in the above step and then put into a four-hole culture plate to be cultured; and step 3: medicine lentiviral, egg pumping liquid, the maturation fluid, micromanipulation liquid, SOF, the fertilization fluid, the culture fluid and sheep serum are prepared.

The invention provides a screening culture method for sheep oocytes in vitro, which comprises the following steps: step 1: ovaries which were killed less than half an hour ago are put into saline water at the temperature of 30DEG C to 40DEG C and containing penicillin and streptomycin, washed for 3 to 4 times within 3 hours, and oocytes are picked out; 25 to 30mu mol / L brilliant cresyl blue is put into 35DEG C to 39DEG C water bath to be dyed for 85 to 92min, the cytoplasm is blue and is washed for 3 to 4 times by in vitro maturation fluid, is put into 55 to 78mul / drip in vitro maturation culture fluid by 25 to 30m / drip, and is cultured in 5 percent CO2 by 95 percent; step 2: cumulus cells are removed from oocytes which are maturated in vitro for 25 to 28 hours; IVF washing liquid is used to wash for 2 to 4 times, and is dripped into 50 to 70mu l fertilization fluid by 25 to 30m / drop; semen is unfrozen, the fertilization fluid is moved in, supernatant fluid is centrifugurated for 4 to5min and removed, the precipitated sperms are added into fertilization fluid drips by the density of 2 to 4*106 / ml and incubated; eggs which are fertilized for 12 to 18 hours are treated in the above step and then moved into a four-hole culture plate to be cultured; and step 3: reagent egg absorption liquid, brilliant cresyl blue maturation liquid, dyeing liquor SOF, the fertilization fluid, the culture fluid and sheep serum are prepared.

A method for detecting the residual amount of Pichia host protein in recombinant human lysozyme raw materials, comprising preparing recombinant human lysozyme, preparing Pichia host protein, detecting the total protein content of Pichia host protein, and preparing Pichia host protein Antiserum, elimination of cross-reaction between recombinant human lysozyme and Pichia host protein immune rabbit serum and Pichia host protein immune sheep serum, two-dimensional electrophoresis of Pichia host protein, detection of antibody coverage of Pichia host protein, enzyme Composition of linked immunoassay steps. The Pichia host protein in the recombinant human lysozyme is detected in three consecutive batches by using the detection method of the present invention, and the content of the Pichia host protein is all <0.05%, meeting the <0.1% standard stipulated in the Pharmacopoeia of the People's Republic of China. The detection method of the invention has the advantages of strong specificity, full coverage of antibodies, no cross-reaction, etc., and can be used for content detection of Pichia pastoris host protein.

The invention discloses a quality control product, comprising a quality control product 1 and a quality control product 2, characterized in that, the quality control product 1 comprises the following components: 2-6 parts by weight of factor-rich sheep plasma, 2-5 parts by weight The goat serum in parts by weight and the fibrinogen mother liquor in 1-4 parts by weight; the quality control product 2 includes the following components: 1-4 parts by weight of factor-deficient goat plasma, 5-10 parts by weight of goat serum and 0.3- 2 parts by weight of fibrinogen mother solution, the fibrinogen mother solution is prepared by dissolving fibrinogen in goat serum, wherein the mass percentage of fibrinogen is 10%; There is no need to extract coagulation factors, the preparation steps are simplified, the operation is easy, the cost is low, the results are accurate, and the stability is good; Replacing the quality control products produced by the listed American Haemoscope company, reducing the cost of use for customers.