80 results about "Streptomycin" patented technology

The invention discloses a culture medium and application thereof, and a method for in-vitro separation culture and adipogenic differentiation of precursor adipocytes in goat muscle, and relates to the technical field of cell culture, the culture medium comprises the following components: 8-12 [mu] g / mL of insulin, 8-12% by volume of fetal calf serum, 95-105 U / mL of penicillin, 95-105 [mu] g / mL of streptomycin and the like. On the basis of the prior art, a proper amount of liquiritin and pilocarpine are mainly added into a culture medium, and synergistic cooperation of the liquiritin and pilocarpine is found, so that the adipogenic differentiation capacity of cells such as mesenchymal stem cells and precursor adipocytes is remarkably improved, and the application range is wide. On the basis, the invention also provides a complete set of perfect in-vitro isolated culture and adipogenic differentiation method of the goat intramuscular precursor adipocytes, and a method and a way are provided for a series of researches developed through the goat primary intramuscular precursor adipocytes in the future.

The invention belongs to the technical field of cell biology, and particularly discloses a culture medium and a culture method of fish primary liver cells. The culture medium of the fish primary liver cells comprises the following components: 90% of an M199 culture medium, 10% of fetal calf serum, 100 U / mL of penicillin, 0.1 mg / mL of streptomycin, 5 ng / mL of EGF, 10 [mu] U / ml of insulin and 10 mM of dexamethasone. Based on the culture medium, an efficient and stable fish liver cell culture method is developed by combining a standardized anesthesia and disinfection process, a precise dissection and liver sampling method, an optimized tissue digestion and cell separation scheme, an efficient cell purification and adherent culture system, a stable passage technology and the like; a standardized hepatocyte line establishment operation process is defined, and the problems that species binding exists in an existing fish liver cell culture method, and a standardized culture process suitable for multiple fish fingerlings is difficult to form are solved.

The invention belongs to the technical field of microbial pharmacy, and discloses a polyketone macrolide compound generated by genetically engineered streptomyces as well as a preparation method and application thereof. The polyketone macrolide compound disclosed by the invention is a cinnamyl streptomycin derivative, has cell proliferation inhibition activity better than that of a natural product Cinnamomycin A-D, can exert anti-tumor activity by inhibiting activity of human exonucleotide pyrophosphatase / phosphodiesterase 1 (ENPP1) and activating an inherent immune pathway, can be prepared into a medicine or a medicinal composition, and can be used for preparing medicines or medicinal compositions. The compound is used for treating tumors and related diseases.

The invention relates to an isolated culture method of functional Chinese softshell turtle ovarian granular cells. The isolated culture method comprises the following steps: S1, collecting ovarian tissues: collecting Chinese softshell turtle ovarian tissues; s2, separating and collecting a granular cell layer; s3, digesting and collecting granular cells; s4, primary culture of granulosa cells: transferring the collected cells into a cell culture bottle containing 4 ml of a complete culture medium for primary culture, wherein the complete culture medium comprises the following main components: a DMEM culture medium, 15% of fetal calf serum, 1% of penicillin streptomycin and 1% of an insulin-transferrin-selenium additive; and S5, subculturing the granular cells. The isolated culture method and the culture medium system provided by the invention are simpler and more time-saving and labor-saving; according to the method, the aromatase CYP19A1 can be expressed normally, the characteristics of the granular cells are kept, the aromatase CYP19A1 can be expressed normally, it is indicated that the in-vitro cultured cells can keep the functionality of the cells, and the in-vitro proliferation capacity of the granular cells is improved to a great extent.

The application discloses a preparation method of a nanogel preparation for preventing and treating tobacco diseases, and specifically comprises the following steps: (1) preparing bacillus licheniformis bacterial liquid; (2) preparing nanometer selenium precipitate; (3) preparing purified nanometer selenium particles; (4) preparing streptomycin-nanometer selenium solid; (5) preparing nanometer titanium dioxide photo-semiconductor sol; and (6) mixing low-melting agarose powder, the streptomycin-nanometer selenium solid and the nanometer titanium dioxide photo-semiconductor sol, centrifuging, washing, and drying to obtain the nanogel preparation. The nanogel preparation has significant bacteriostatic efficacy, can effectively prevent and treat tobacco diseases (especially tobacco bacterial wilt and tobacco wild fire disease), has a promoting effect on tobacco growth, is simple to make, has strong adhesion, and does not pollute the soil environment.

The invention discloses an amplification culture solution of TILs cells and a culture method thereof. The amplification culture solution comprises a complete culture medium, and an anti-BTLA antibody with the final concentration of 0.1-10 [mu] g / mL, penicillin with the final concentration of 50-500 [mu] g / mL, streptomycin with the final concentration of 50-500 [mu] g / mL, gentamicin with the final concentration of 10-100 [mu] g / mL and amphotericin with the final concentration of 1-50 [mu] g / mL are added into the complete culture medium. The anti-BTLA antibody component in the culture solution can relieve inhibition of BTLA on T cells by blocking combination of BTLA and a ligand HVEM of BTLA, so that the anti-tumor capability of TILs is enhanced; meanwhile, after the BTLA is combined with the ligand HVEM, T cell activation and proliferation can be inhibited, and rapid proliferation of TILs is promoted; the irradiation PBMC added in the TILs cell culture process can provide an activation signal for the TILs and drive rapid proliferation of the TILs cells.

The present invention belongs to the technical field of sensorineural hearing. Provided is the use of SPTBN1 as a target in the treatment of GJB2-related sensorineural hearing loss. The use comprises: the construction of a stably transfected cell line, wherein: HEK293T cells are cultured using a DMEM culture medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin in a humidified incubator at 37ºC with 95% air and 5% CO2, and when the cells reach 80%-90% confluence, cell passage is performed; IP-MS analysis, wherein: protein complexes are purified using Protein A / G immunoprecipitation magnetic beads, a portion of the extracted proteins is used as input, and then 2 μg of anti-Cx26 antibody is added to the remaining protein extract, same are gently pipetted and mixed, and incubated on a rotating shaker at 4℃ overnight; immunofluorescence observation of the co-localization of Cx26 and SPTBN1, wherein: a stably transfected cell line expressing WT-Cx26 and Mut-Cx26 is constructed; and co-immunoprecipitation (Co-IP) validation, wherein: a stably transfected cell line expressing WT-Cx26 is constructed. The present invention overcomes the limitations of therapeutic methods, such as the relatively low targeting specificity and short therapeutic time windows associated with full-length protein supplementation via gene therapy, thereby providing new insight into GJB2-related hearing loss, and a new target and a new treatment for same.

The invention belongs to the field of biological medicine, and relates to a curcumin-loaded bone marrow mesenchymal stem cell source exosome which is obtained by the following steps: step 1, culturing BMSCs (bone marrow mesenchymal stem cells): simultaneously adding 10% of fetal calf serum and 1% of myclostreptomycin into an MEM culture medium, culturing at 37 DEG C under 5% CO2, and counting and subculturing when the cell density reaches 90%; 2, dissolving Cur in a DMSO solution, preparing a mother solution with the culture medium, and filtering and diluting to 30-50 [mu] g / mL; adding into BMSCs (bone marrow mesenchymal stem cells) with the cell density of 85-100%, and culturing for 12-36 hours; collecting supernate, and carrying out gradient centrifugation to obtain the mesenchymal stem cell source exosome loaded with curcumin. The invention provides a novel administration route which depends on the self function of the mesenchymal stem cell source exosome and the self function of the curcumin at the same time.

The invention belongs to the technical field of nerve cell culture, and particularly relates to application of a culture medium in co-culture of SVGP12 and HMEC-1. The culture medium is based on a mixture of DMEM and MCDB 131 according to a ratio of 1: 1, HEPES, Glutamax, N2, G-5, B-27, hEGF, penicillin-streptomycin and fetal calf serum are added, and the components are prepared according to a specific final concentration. When being applied to a Transwell co-culture model, the culture medium can meet the nutritional requirements of two cells at the same time, maintain the normal growth and form of the cells, and promote the transmission of functional signal molecules. Experiments show that the culture medium can significantly up-regulate expression of ABCG2, CYP2D6, CYP1B1 and CYP2E1 in HMEC-1 (P is less than 0.05), provides a reliable in-vitro model for researching an interaction mechanism of two cells, and assists drug screening and disease simulation.

The application belongs to the technical field of microorganism detection, and particularly relates to a bifidobacterium viable count detection culture medium, a detection method and application. The application provides a culture medium for detecting bifidobacterium, wherein the culture medium comprises MRS culture medium and an antibiotic solution; the antibiotic solution comprises streptomycin, kanamycin and sodium propionate. The culture medium can ensure normal growth of bifidobacterium, has strong inhibition on lactobacillus, can effectively realize accurate detection of bifidobacterium, and avoids interference of lactobacillus.

The application provides a preparation method of a ratio fluorescent aptamer sensor for detecting streptomycin in milk, and belongs to the field of food safety detection. The application comprises screening of streptomycin (STR) specific aptamer and construction of a ratio fluorescent sensor. The STR aptamer (Apt) is obtained by using a magnetic graphene oxide screening technology for the first time, the binding site is analyzed according to the secondary structure and molecular docking analysis, and the Apt52 with the best affinity to the target is obtained by truncation. The carbon quantum dots (CQDs) modified by the Apt52 are used as a reference signal, the AgNCs-SMP@ZIF-8 is used as a response signal to construct a fluorescent sensing method for judging whether the streptomycin is contained in the sample to be detected. The specific binding of the aptamer to the target causes the AgNCs-SMP@ZIF-8 to expose and quickly adsorb a large amount of Cu 2+ The fluorescence signal of the AgNCs-SMP@ZIF-8 is quenched, and the fluorescence signal of the CQDs always remains stable, and the streptomycin residue is detected by the fluorescence signal ratio change of the AgNCs-SMP@ZIF-8 and the CQDs. The detection limit of the sensor for the STR is 0.978 nM, and the sensor has satisfactory specificity and sensitivity.

This invention relates to the field of gastric cancer organoid microarray technology, and more particularly to a gastric cancer organoid microarray culture medium and its applications. The invention provides a novel culture medium adapted for gastric cancer organoid microarray culture. The medium components include basal medium, penicillin / streptomycin dual antibiotics, antibacterial agents, hydroxyethylpiperazine ethanesulfonic acid, B-27 additive, Glutamax culture medium additive, R-vertebral protein 1, scutellarin B, and tetramethylpyrazine, among other components. This invention solves key problems in existing technologies such as slow growth, morphological instability, poor heterogeneity adaptation, and low long-term culture activity of gastric cancer organoids, providing an ideal experimental model for high-throughput screening and precision medicine applications on microarrays.

The invention belongs to the technical field of nerve cell culture, and particularly relates to a co-culture method of SVGP12 and HMEC-1 cells. The method comprises the following steps: firstly, preparing a co-culture medium, on the basis of mixing DMEM and MCDB 131 according to a volume ratio, adding HEPES, Glutamax, N2, G-5, B-27, hEGF, penicillin-streptomycin and fetal calf serum, respectively adopting products with corresponding goods numbers for each component, and controlling specific final concentrations; and then respectively inoculating SVGP12 and HMEC-1 into culture flasks, culturing by using the co-culture medium, and carrying out passage. The method can realize indirect communication of two cells in a shared microenvironment, provides a reliable tool for researching a regulation mechanism of SVGP12 on HMEC-1, and also provides a reference for optimization design of a co-culture system.

The invention provides Kineococcus G2 and application of the Kineococcus G2 in preparation of a pigment with antioxidant activity, the Kineococcus new strain G2 is CCTCC No. M20242602, the strain is spherical, the size is 1.0 * 1.5 mu m, the optimal growth temperature is 30-37 DEG C, the pH value is 7.0-9.0, and the salt concentration is 1.0%-2.0%. The strain G2 is not sensitive to bacitracin, ciprofloxacin and norfloxacin, and is sensitive to novobiocin, polymyxin, gentamicin, piperacillin, ofloxacin, erythromycin, streptomycin, kanamycin, rifampicin, ampicillin, vancomycin, carbenicillin, chloramphenicol, tetracycline, penicillin, cefoperazone, oxacillin, amoxicillin and neomycin. Pigment generated by fermentation of the G2 strain has good antioxidant activity, and when the concentration is 10 micrograms / mL, the scavenging activity on DPPH is 25.00%, the scavenging activity on hydroxyl radicals is 80.58%, and the iron ion reducing capacity is 1.57, which are all higher than those of contrast beta-carotenoid. The application of the new Kineococcus strain G2 can lay a foundation for the research of antioxidant drugs such as medical treatment and health care product development.

The present disclosure relates generally to methods and systems for controlling Candidatus Liberobacter Disease in citrus plants, including, for example, orange trees. In some embodiments, the methods comprise injecting an injection formulation comprising a combination of oxytetracycline and streptomycin into a tree.

The separation and detection method comprises the following steps: rinsing placenta or villus tissues in a PBS (Phosphate Buffer Solution) added with a 1% penicillin-streptomycin double antibody solution, digesting the placenta or villus tissues into single cells by using a 1640 culture medium containing I-type and IV-type collagenase after the placenta or villus tissues are rinsed sufficiently to obtain a cell suspension, and culturing the cell suspension for later use; diluting and neutralizing collagenase digestive juice by using a 1640 culture medium containing high-temperature heat-inactivated fetal calf serum according to 5 times of volume; repeatedly discarding the precipitate, recovering the supernatant, and centrifuging to obtain a coarse migration body; and establishing a density gradient by using Optiprep as a density medium, washing the precipitate with PBS, and centrifuging to obtain the trophoblast cell-derived transporter. The invention provides a group of placenta villus trophoblast cell source migration body specific expression membrane proteins for the first time, and provides a method for specifically capturing the placenta trophoblast source migration body on the basis of the placenta villus trophoblast cell source migration body specific expression membrane proteins, so that a new direction for researching a trophoblast dysfunction related disease mechanism can be enriched.

This invention proposes a method for preparing dental pulp stem cell exosomes and their applications, belonging to the field of exosome technology. Dental pulp stem cells from exfoliated deciduous teeth are seeded into serum-free α-MEM medium containing microparticles, insulin-like growth factor, transforming growth factor-β1, penicillin, and streptomycin for culture. Exosomes are isolated, dispersed in PBS solution, and incubated with microparticles, propylene glycol, and rhodioloside to obtain highly active dental pulp stem cell exosomes. The surface of these exosomes is then modified with shikonin and interferon to obtain dental pulp stem cell exosomes. The dental pulp stem cell exosomes prepared by this invention exhibit good affinity for titanium implant-mediated osteointegration repair materials, demonstrating good anti-inflammatory, immunomodulatory, osteogenic, and damage-reducing effects. Furthermore, the preparation method is simple, yields high output, and operates under mild conditions, showing broad application prospects.

The application discloses a method and a kit for immunodetection of target substances by using arterial blood, and belongs to the technical field of blood detection. The kit comprises at least: a first component containing luminescent microspheres combined with detection antibody 1 and a luminescent microsphere diluent, wherein the luminescent microsphere diluent contains 2-morpholinoethanesulfonic acid, KCl and NaHCO3; a second component containing biotin-labeled detection antibody 2 and a biotin diluent, wherein the biotin diluent contains dithiothreitol, CaCl2, adenine nucleoside and mouse anti-human hemoglobin antibody; and a third component containing photosensitive microspheres combined with streptomycin. The scheme can be used for immunodetection of target substances by using arterial blood, has excellent functional sensitivity and detection range, is not interfered by other substances in arterial blood, realizes rapid detection of all blood gas indexes and immunological indexes by using one tube of arterial blood, and has important significance for clinical blood detection.

The invention discloses a skincare stem cell exosome separation and extraction method, and relates to the technical field of biological medicine, the skincare stem cell exosome separation and extraction method comprises the following steps: S1, culture of mixed stem cells: unfreezing urine-derived stem cells and pilose antler stem cells, resuspending in an improved DMEM culture medium, and carrying out mixed culture to prepare a mixed culture solution; epidermal growth factors, cholera toxin, penicillin, streptomycin, transferrin, IL-2 and IL-15 are added into the improved DMEM culture medium; s2, pretreatment of the mixed culture solution: performing centrifugal treatment on the mixed culture solution, and separating to obtain a precipitate and a supernatant, the precipitate being mixed stem cells. The urine-derived stem cells and the pilose antler stem cells are subjected to mixed culture and matched with the improved culture medium containing IL-2, IL-15 and other components, so that the yield and biological activity of the exosome are remarkably improved, high purity and low impurity protein residue of the exosome are achieved by means of a complete purification process of multi-step centrifugation, ultrafiltration concentration and filter membrane filtration, and the yield of the exosome is improved. The skin allergy stimulation risk is effectively reduced.

The invention belongs to the technical field of cell culture, and particularly relates to a culture medium special for HMEC-1 cells, application and a culture method. The special culture medium is prepared from the following components in percentage by volume: 10% of fetal calf serum (FBS), 89% of DMEM basal culture medium and 1% of penicillin-streptomycin double-antibody solution (P / S). According to the special culture medium and the culture method, efficient proliferation and stable culture of the HMEC-1 cells can be guaranteed, expensive components do not need to be additionally added, the cost is controllable, and a reliable culture scheme is provided for related research of the HMEC-1 cells.

The invention provides a dental pulp stem cell exosome as well as a preparation method and application thereof, and belongs to the technical field of exosomes. The method comprises the following steps: inoculating exfoliated deciduous tooth pulp stem cells into a serum-free alpha-MEM culture medium containing microparticles, insulin-like growth factors, transforming growth factor-beta1, penicillin and streptomycin for culturing, separating exosomes, dispersing the exosomes into a PBS solution, adding microparticles, propylene glycol and salidroside for incubation, and separating to obtain the high-activity pulp stem cell exosomes. And performing surface modification on alkannic acid and interferon to prepare the dental pulp stem cell exosome. The dental pulp stem cell exosome prepared by the invention has relatively good affinity to a titanium implant material mediated osseointegration repair material, has relatively good advantages of resisting inflammation, regulating immunity, promoting osteogenesis, reducing injury and the like, is simple in preparation method, relatively high in yield and mild in condition, and has a wide application prospect.

The application discloses a culture medium and application thereof, and a method for in-vitro separation and culture of goat intramuscular precursor fat cells and adipogenic differentiation of the cells, and relates to the technical field of cell culture. The culture medium comprises the following components: 8-12 mu g / mL insulin, 8-12% fetal bovine serum in volume fraction, 95-105 U / mL penicillin, 95-105 mu g / mL streptomycin, and the like. On the basis of the prior art, the application mainly adds appropriate glycyrrhizin and pilocarpine in the culture medium, and finds that the two components are in cooperation, and the adipogenic differentiation ability of cells such as mesenchymal stem cells and precursor fat cells is significantly improved, and the application range is wide. On the basis, the application further provides a complete method for in-vitro separation and culture of goat intramuscular precursor fat cells and adipogenic differentiation of the cells, and provides a method and a way for a series of researches on goat primary intramuscular precursor fat cells in the future.

The invention discloses a method for in-vitro broad-spectrum stimulation of immune checkpoint up-regulation and application, the method comprises the following steps: placing a Jurkat cell line in an RPMI-1640 culture medium containing 10% of fetal calf serum and 1% of penicillin-streptomycin, carrying out in-vitro stimulation for 24-72 hours by adopting PHA with a concentration of 0.1-5 [mu] g / mL under the conditions of 37 DEG C and 5% of CO2, activating NF-kB and / or MAPK signal channels, and carrying out in-vitro broad-spectrum stimulation of immune checkpoint up-regulation. The broad-spectrum up-regulation of co-stimulatory immune checkpoints of CD28, CD40L, ICOS and the like and co-inhibitory immune checkpoints of PD-1, LAG-3, CTLA-4 and the like is realized, and the high expression state can be maintained for at least 72 hours after the PHA is removed. The invention also discloses the in-vitro broad-spectrum immune checkpoint positive T cell model constructed by the method, and the up-regulation of immune checkpoints in the model is accompanied by the increase of dose-dependent secretion of various cytokines. The model can replace a traditional model, is applied to toxicology and pharmacology research of immune checkpoint antagonists and combined synergistic effect and immune checkpoint regulation and control mechanism research, is simple and convenient to operate and high in repeatability, and provides an efficient tool for research and development of related drugs.

The invention discloses a phosphorus-solubilizing fungus obtaining and optimizing method, and belongs to the field of environment and the field of agricultural biotechnology. The method comprises the following steps: collecting a soil sample of Inner Mongolia Tin-Go Union; performing liquid culture on the soil sample, and performing gradient dilution under a sterile condition; the method comprises the following steps: sucking 100 [mu] L of bacterial solutions with different concentrations, coating the bacterial solutions on a PVK solid culture medium added with chloramphenicol and streptomycin, culturing, selecting single colonies with good growth, typical characteristics and large and remarkable phosphate solubilizing circle, continuously carrying out streak purification on the PVK solid culture medium to obtain a target strain pure culture, preserving and identifying to obtain the phosphate solubilizing fungi; optimizing culture conditions: taking glucose as a unique carbon source, wherein the concentration of the glucose is 15g / L; ammonium chloride is used as a sole nitrogen source, and the concentration of the ammonium chloride is 0.082 g / L; the culture temperature is 30 DEG C; the initial pH is 5; the phosphorus solubilizing amount of the phosphorus solubilizing fungi is 858.31 mg / L.

The invention discloses a thyroid cancer organoid culture medium, a thyroid cancer organoid culture method and a drug sensitivity detection method, and belongs to the technical field of organoid models. The thyroid cancer organoid culture medium provided by the invention comprises a basic culture medium and the following components: 0.5 to 2 percent (V / V) of N2, 0.5 to 2 percent (V / V) of penicillin-streptomycin-amphotericin, 0.5 to 5 mM of N-acetylcysteine, 5 to 30 mM of Nicotinamide, 1 to 10 mu M of CHIR99021, 5 to 50 ng / mL of hEGF, 1 to 20 mu M of a TGF-beta receptor inhibitor and 1 to 50 M of a p38 MAPK inhibitor. The thyroid cancer organoid culture medium is utilized in the thyroid cancer organoid culture process, the improvement of a culture method and a drug sensitivity detection method is combined, the organoid construction efficiency reaches up to 90% or above, the thyroid cancer organoid culture and drug sensitivity period can be greatly shortened, and the thyroid cancer organoid culture medium is low in cost and easy to popularize. Good popularization and application prospects are realized.

This invention discloses a method for microscopically separating and rescuing cells from heavily contaminated cell culture media, belonging to the field of biotechnology. This invention combines flow cytometry sorting technology with penicillin-streptomycin for the separation, rescue, and culture of healthy cells. The method provided by this invention can effectively rescue heavily contaminated cells, remove their contamination state, and make them usable again. This will greatly facilitate cell preservation, passage, and culture, effectively avoid the waste of cell resources, and improve the preservation rate of rare cell resources, providing strong support for the smooth progress of scientific research.

The invention provides an early-stage chick embryo blood collection method and application thereof, and belongs to the technical field of poultry genetic engineering. The method for collecting the blood of the early chick embryo comprises the following steps: hatching a fertilized hatching egg for 56-60 hours, enabling the tip of the hatching egg to face upwards, windowing at the middle position between the tip and the equator end, shaking to observe the chick embryo, sucking the blood in the dorsal aorta blood vessel of the chick embryo by adopting a mouth suction tube method, then adding a penicillin streptomycin solution, sealing, and continuing hatching. According to the method for collecting the blood of the early chick embryos, the subsequent survival and development of the chick embryos are not affected, the survival rate of the chick embryos subjected to blood collection through the method is high, the chick embryos can be successfully hatched, and the hatching rate is similar to that of wild types without blood collection. The early chick embryo sex identification method provided by the invention can directly identify the sex of the chick embryo through trace blood, is simple and easy to implement, does not influence the subsequent survival and development of the chick embryo, and has the advantages of high efficiency, rapidness, accuracy and high sensitivity.

The invention discloses a method for detecting influence of bacteria on tumor cell proliferation based on CCK8, which is characterized by comprising the following steps: adding a trigeminy antibiotic culture medium containing penicillin-streptomycin-gentamicin into a bacteria and cell co-culture system to treat the co-culture system; comprising the following steps: centrifuging a cell suspension, discarding a supernatant, adding an antibiotic-free DMEM culture medium, uniformly mixing, and adding cells into a 96-well plate for overnight culture; centrifuging fresh bacterial liquid, discarding supernatant, adding a DMEM (Dulbecco Modified Eagle Medium) without antibiotics, uniformly mixing, adding bacteria into a 96-well plate, culturing, discarding an original culture medium, adding a DMEM containing penicillin-streptomycin-gentamicin antibiotics, and culturing; the method comprises the following steps of: sucking and abandoning a culture medium in a 96-well plate, adding a DMEM (Dulbecco Modified Eagle Medium) containing 10vt% of CCK8 reagent, measuring an absorbance value by using a microplate reader, and judging the influence of bacteria on tumor cell proliferation. The method has the advantages of higher stability and repeatability, and more reliable experimental result.

The application discloses a kind of Vibration Mountain beard toad hind leg / toe tissue cells and its quick acquisition method and application.The method comprises: the body surface disinfection of Vibration Mountain beard individual, cut and take hind leg or toe tissue, and put back after wound disinfection;After tissue is soaked in 75% alcohol and washed with PBS containing penicillin, streptomycin and amphotericin B, it is temporarily stored in 4 ℃ temporary storage solution;After tissue is cut, it is sequentially digested with trypsin and type I collagenase;In the optimized low permeability medium, primary culture is carried out at 25-27 ℃;When cell confluence reaches 60%-70%, it is passaged using trypsin-EDTA digestion;Freezing and recovery.The application realizes the non-lethal cell of Vibration Mountain beard for the first time Quick acquisition, cell first time migration time is only 3 days, grow into monolayer only 12-17 days, pollution rate is reduced to 5.6%, recovery survival rate reaches 76.19%, and provides efficient and reliable technical support for endangered amphibian germplasm resource preservation and biological research.

This invention relates to the field of immunology, and more particularly to a hybridoma cell line and its secreted monoclonal antibody against the metamizole residue marker 4-methylaminoantipyrine (MAA), and its applications. This invention uses 4-aminoantipyrine to react with methyl 4-bromomethylbenzoate, followed by alkaline hydrolysis to obtain a hapten (Formula I). ​​An immunogen is then synthesized and used to immunize mice. After cell fusion and screening, a hybridoma cell line capable of secreting a monoclonal antibody recognizing MAA is prepared, named Metamizole-6A4. This hybridoma cell line has a high efficiency in secreting monoclonal antibodies. The secreted monoclonal antibody can accurately detect MAA, IC50, and other markers. 50 The concentration is 4.06 ng / mL, with high specificity. Cross-reactivity with aminopyrine and its metabolites 4-aminoantipyrine, 4-formamidoantipyrine, 4-acetamidoantipyrine, sulfadiazine, norfloxacin, lincomycin, tylosin, penicillin G, streptomycin, and gentamicin is less than 5%. It can accurately detect MAA residues in food. Formula I.