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300 results about "Western blot" patented technology

<ul><li>A positive test indicates presence of antibody/other specific protein and a negative test indicates its absence.</li><li>In case of infections, a positive test is commonly a result of disease. However, in certain conditions such as agammaglobulinemia where Western blotting is performed to detect normal proteins (BTK protein), absence could be the abnormal result.</li><li>The doctor reviews the results and discusses it.</li></ul>

Monoclonal antibody BY017 combined with African swine fever virus P30 protein and application thereof

The invention discloses a monoclonal antibody BY017 combined with African swine fever virus P30 protein and application of the monoclonal antibody BY017. The invention provides an IgG antibody which is composed of a light chain and a heavy chain. CDR1, CDR2 and CDR3 in the heavy chain variable region are sequentially shown as the 47 to 54 , the 72 to 79 and the 118 to 126 in SEQ ID NO: 1; cDR1, CDR2 and CDR3 in a light chain variable region are sequentially as shown in the 49 to 60 , the 78 to 80 and the 117 to 124 in SEQ ID NO: 3. The invention also protects the application of any one of the IgG antibodies in preparation of products. The product has the functions that the product is used for being combined with an African swine fever virus P30 protein fragment; the antibody is used for binding African swine fever virus P30 protein; the antibody is used for binding African swine fever virus. The IgG antibody provided by the invention can be used for multiple purposes of western blot, immunohistochemistry, flow cytometry, ELISA (Enzyme-Linked Immunosorbent Assay) and the like, and can be used as an important tool for ASFV detection and structural analysis of P30 protein of ASFV detection.
Owner:베이징 중커 란위 바이오테크놀로지 씨오 엘티디 +1

Antibody aiming at severe fever with thrombocytopenia syndrome virus glycoprotein and application thereof

The monoclonal antibody of the glycoprotein Gn of the severe fever with thrombocytopenia syndrome virus is screened on the basis of immune repertoire sequencing, and the biological characteristics of the monoclonal antibody are identified. The method comprises the following steps: taking purified SFTSV-Gn as bait, taking virus specific B cells from an immunized female BALB / c mouse B cell bank, selecting an antibody sequence with potential binding activity by utilizing a 10 * immune repertoire sequencing method in combination with bioinformatics analysis, constructing the antibody sequence to an eukaryotic expression vector pcDNA3.4, expressing and purifying a monoclonal antibody, and identifying the biological activity of the monoclonal antibody. 14 antibodies are selected from 3544 captured B cells, functional verification shows that 12 antibodies can be combined with SFTSV-Gn glycoprotein, and the Gn-5 antibody and the Gn-7 antibody have high combining capacity; western blot, indirect immunofluorescence test and flow cytometry prove that the antibody can specifically recognize the SFTSV, the effective concentration is as low as 2 mu g / mL, and a foundation is laid for development of a rapid diagnosis reagent for the SFTSV.
Owner:SHANDONG FIRST MEDICAL UNIV & SHANDONG ACADEMY OF MEDICAL SCI

Anti-CD3D antibody and application thereof

The invention discloses an anti-CD3D antibody and application thereof.The antibody comprises a heavy chain variable region and a light chain variable region, the amino acid sequence of CDR-H1 of the heavy chain variable region is shown as SEQ ID NO: 4, the amino acid sequence of CDR-H2 of the heavy chain variable region is shown as SEQ ID NO: 5, and the amino acid sequence of CDR-H3 of the heavy chain variable region is GDL; the amino acid sequence of the CDR-L1 of the light chain variable region is as shown in SEQ ID NO: 6, the amino acid sequence of the CDR-L2 of the light chain variable region is as shown in SEQ ID NO: 7, and the amino acid sequence of the CDR-L3 of the light chain variable region is as shown in SEQ ID NO: 8. The anti-CD3D antibody is a rabbit monoclonal antibody, the obtained antibody is high in titer, clear in specific positioning in immunohistochemical staining and free of non-specific background staining, the antibody is wide in application, and the antibody can be used in pathological immunohistochemical staining experiments and can also be used in Western blot and flow cytometry experiments.
Owner:HANGZHOU BIOLYNX TECH CO LTD

Western blot image generation forgery detection method and system, terminal and storage medium

The invention belongs to the technical field of image processing, and discloses a western blot image generation forgery detection method and system, a terminal and a storage medium, and the method comprises the steps: obtaining a to-be-detected image; wherein the to-be-detected image comprises a face image and a western blot image; performing feature extraction on the to-be-detected image based on a counterfeit detection model to obtain visual features; performing generation discrimination on the visual features to obtain a real discrimination result or a counterfeit discrimination result of the to-be-detected image; source discrimination is carried out on the visual features corresponding to the counterfeit discrimination result, and a discrimination result that the to-be-detected image comes from the face image or the western blot image is obtained; and outputting the western blot image to generate a forgery detection result. According to the method, the forgery recognition precision of the western blot image is improved, and the generalization ability of image forgery detection is enhanced.
Owner:SHENZHEN MSU-BIT UNIVERSITY +1

Construction method and application of polycystic ovarian syndrome endometrial epithelium organ fibrosis model

The invention relates to the technical field of biology, and particularly discloses a construction method and application of an endometrial epithelium organ fibrosis model. According to the method, human endometrial epithelial cells are extracted, and an endometrial epithelial organ is established by adopting a Matrigel three-dimensional culture system. Afterwards, the organoid is induced to form endometrial phenotypes in different physiological cycles through hormone stimulation, and a ferroptosis inducer FIN56 is added in the culture process to induce fibrosis. The expression of GPX4, Fibronectin and p-Smad2 / 3 is detected through Western blot and immunofluorescence, and the successful construction of the fibrosis model is verified. The invention provides an in-vitro model capable of simulating endometrial fibrosis pathological characteristics of a patient with PCOS (polycystic ovarian syndrome), the in-vitro model can be used for mechanism research and drug screening of PCOS-related endometrial lesions, and a new research tool and an intervention strategy are provided for treatment of PCOS endometrial fibrosis.
Owner:PEKING UNIVERSITY THIRD HOSPITAL (THE THIRD CLINICAL MEDICAL SCHOOL OF PEKING UNIVERSITY)

Method for detecting activity of HPV (human papillomavirus) trivalent therapeutic vaccine

The invention relates to the technical field of vaccines, in particular to an activity detection method of an HPV (human papilloma virus) trivalent therapeutic vaccine. The method comprises the following steps: inoculating and infecting BEAS-2B cells by adopting constructed HPV16 / 18 / 52 type E6 / E7 antigen-loaded non-replicating type 5 adenovirus, culturing, extracting total protein, sequentially performing SDS-PAGE (Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis), western blot detection, immunoreaction and developing imaging, and detecting the fusion protein; the transfection complex number MOI of the constructed adenovirus is 15-25, and the convergence degree of BEAS-2B cells is greater than 80%; the membrane transfer current of the western blot method is 220-280 mA, and the time is 50-80 minutes. The method avoids the problem of inaccurate detection results caused by obvious difference of biological activities in different animal species, has good accuracy, stability and durability, is simple in steps and low in cost, and can be effectively popularized and used.
Owner:TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH

Insulin resistance method for promoting signal pathway based on CTRP9

The invention relates to the technical field of insulin resistance, in particular to an insulin resistance method for promoting a signal channel based on CTRP9. The method comprises the following steps: constructing a human hepatocyte line plasmid; carrying out cell culture and transfection; detecting the influence of CTRP9 on LO human hepatocyte glucose uptake, PGC-1a gene expression and the phosphorylation level of insulin signal channel related protein; detecting the influence of CTRP9 on glucose uptake of an LO human hepatocyte insulin resistance model, PGC-1a gene expression and the phosphorylation level of insulin signal channel related protein; and carrying out SirT1 / PGC-1a protein expression detection. According to the insulin resistance method for promoting the signal channel based on the CTRP9, a human hepatocyte line is taken as an in-vitro research object, an in-vivo research environment is simulated, a corresponding insulin resistance model is established, and the insulin resistance can be obtained by using technologies such as flow cytometry analysis, RT-PCR (Reverse Transcription-Polymerase Chain Reaction), western blotting and signal channel inhibition. And the treatment effect of CTRP9 on hepatic cell insulin resistance and the specific action mechanism are researched.
Owner:CHONGQING NO 9 PEOPLES HOSPITAL

Monoclonal antibody BY013 combined with African swine fever virus P30 protein and application thereof

The invention discloses a monoclonal antibody BY013 combined with African swine fever virus P30 protein and application of the monoclonal antibody BY013. The invention provides an IgG antibody which is composed of a light chain and a heavy chain. CDR1, CDR2 and CDR3 in the heavy chain variable region are sequentially shown as the 47 to 54 , the 72 to 79 and the 118 to 130 in SEQ ID NO: 1; cDR1, CDR2 and CDR3 in a light chain variable region are sequentially as shown in the 49 to 60 , the 78 to 80 and the 117 to 124 in SEQ ID NO: 3. The invention also protects the application of any one of the IgG antibodies in preparation of products. The product has the functions that the product is used for being combined with an African swine fever virus P30 protein fragment; the antibody is used for binding African swine fever virus P30 protein; the antibody is used for binding African swine fever virus. The IgG antibody provided by the invention can be used for multiple purposes of western blot, immunohistochemistry, flow cytometry, ELISA (Enzyme-Linked Immunosorbent Assay) and the like, and can be used as an important tool for ASFV detection and structural analysis of P30 protein of ASFV detection.
Owner:베이징 중커 란위 바이오테크놀로지 씨오 엘티디 +1

Application of THBS4 inhibitor in preparation of medicine for preventing and / or treating pulmonary arterial hypertension

The invention relates to an application of a THBS4 inhibitor in preparation of a medicine for preventing and / or treating pulmonary arterial hypertension. Three pulmonary arterial hypertension rat models (a hypoxia induction model, a hypoxia combined SUGEN induction model and a monocrotaline induction model) are constructed, the expression change of THBS4 is verified through qRT-PCR and western blot, and the correlation between the expression level of THBS4 and pulmonary arterial hypertension is verified in combination with in-vitro cell experiments and in-vivo intervention. Results show that THBS4 is remarkably up-regulated in pulmonary arteries of the three models, the expression level of THBS4 is positively correlated with the severity of diseases, and THBS4 is an important regulatory factor for promoting vascular remodeling in pulmonary arterial hypertension. In vivo, by knocking down THBS4, pulmonary vascular remodeling and right ventricular hypertrophy are remarkably relieved, and it is prompted that THBS4 inhibitors show good application prospects in the aspect of preventing and / or treating pulmonary arterial hypertension.
Owner:SHENZHEN UNIV

Construction and verification method of HEK293T cell line for stable overexpression of human ANT2

PendingCN121380126AFermentationGenetic engineeringHuman cloningWestern blot
The invention discloses a construction and verification method of an HEK293T cell line capable of stably overexpressing human ANT2, which comprises the following steps: cloning a human ANT2 gene, constructing the human ANT2 gene into a lentivirus expression plasmid pLVX-TRE3G, co-transfecting the lentivirus expression plasmid pLVX-TRE3G and a helper plasmid pLVX-Tet3G into an HEK293T-ACE2 cell, and carrying out puromycin and G418 pressurized screening and Western blot identification to obtain the HEK293T cell line capable of stably overexpressing human ANT2. A result shows that the expression quantity of the ANT2 is the highest when the HEK293T-ACE2-ANT2 cell is induced for 12 hours at the Doxycycline of 6mg / L; when the ANT2 is induced to express, the virus nucleic acid load and the N protein expression level after SARS-CoV-2 infection are obviously reduced, which indicates that the ANT2 has an inhibition effect on SARS-CoV-2 infection replication, and a cell model and an experimental basis are provided for exploring the effect of the ANT2 in virus infection resistance.
Owner:ACAD OF MILITARY SCI PLA CHINA ACAD OF MILITARY MEDICAL SCI INST OF MILITARY VETERINARY MEDICINE

Epidermal fibroin (FLG) gene and protein detection method based on oral cell non-invasive sampling

The invention discloses an FLG gene and protein detection method based on oral cell non-invasive sampling, which comprises the following steps: Step 1: sample collection: collecting oral cells through mouth rinsing of a subject; step2: RNA extraction and gene expression analysis: extracting oral cell total RNA in the oral cells in the step 1 by using a Trizol reagent, and analyzing FLG gene expression through qRT-PCR (Quantitative Reverse Transcription-Polymerase Chain Reaction); 3, protein extraction and immunoblotting analysis are conducted, specifically, protein is extracted from the oral cells in the step 1 through an SDS lysis buffer solution, and FLG protein expression is detected through the Western blot technology. According to the present invention, the subject rinses the mouth and collects the oral cavity cells, such that the convenient, non-invasive and efficient sample collection is achieved.
Owner:WENZHOU-KEAN UNIV

Immunofluorescence detection method and system for dynamically detecting mouse uterus structure development based on multiple time points

The invention provides an immunofluorescence detection method and system for dynamically detecting mouse uterus structure development based on multiple time points, and immunofluorescence staining comprises the steps of sheet unfolding, dewaxing, repairing, cell membrane punching, sealing, primary antibody treatment and secondary antibody treatment. The system comprises a sample pretreatment control module, a multi-channel confocal microscope imaging module and a data analysis module integrating fluorescence intensity calculation, Pearson co-localization analysis and a Western blot data integration algorithm. Through a multi-time-point and multi-index immunofluorescence detection method, dynamic and systematic data support is provided for mouse uterus development mechanism research, and an application basis is provided for fundamental research of reproductive biology and construction of related disease models.
Owner:GUANGDONG MEDICAL UNIV

CXCL6-targeting blocking antibody and application thereof in preparation of antitumor drugs

The invention relates to the technical field of biological medicines, and discloses a blocking antibody targeting CXCL6 and an application of the blocking antibody in preparation of antitumor drugs. The antibody or antigen-binding fragment thereof is capable of binding specifically to human CXCL6 protein with high affinity (e.g., Kd < = 1 nM). According to the invention, the specificity and high affinity of the antibody are verified by Western Blot, immunofluorescence, immunohistochemistry and surface plasmon resonance (SPR) technologies. Functional experiments show that the antibody can effectively block CXCL6-induced neutrophil chemotaxis, including in Transwell, a three-dimensional gel model and a zebra fish living body model. Besides, in a mouse MC38 colon cancer model, the antibody shows remarkable anti-tumor activity and can inhibit tumor growth driven by CXCL6. The invention also provides a pharmaceutical composition containing the antibody, and application of the antibody in preparation of drugs for preventing or treating diseases (such as inflammatory diseases, autoimmune diseases and cancers) related to abnormal expression or activity of CXCL6.
Owner:AFFILIATED HOSPITAL OF JINING MEDICAL UNIV

Programmed death receptor ligand 2 monoclonal antibody 8e3 and uses thereof

The present application relates to the technical field of biotechnology, in particular to a programmed death receptor ligand 2 monoclonal antibody and application thereof. The present application provides a PD-L2 monoclonal antibody, amino acid sequences of three CDR regions of a heavy chain thereof; amino acid sequences of three CDR regions of a light chain thereof; or amino acid sequences obtained by substitution, deletion or addition of one or more amino acids of the amino acid sequences, or amino acid sequences functionally identical or similar to the amino acid sequences; or amino acid sequences having at least 80% homology with the sequences. The present application widens the possibility of antibody therapy for blocking PD1 / PD1 ligand, and compared with existing PD-L2 monoclonal antibodies, the 8E3 antibody has unique amino acid sequences and CDR region sequences, high affinity and high specificity. The 8E3 antibody can be used in ELISA, Western blot and flow cytometry detection, and has wide application.
Owner:SHENZHEN INST OF ADVANCED TECH CHINESE ACAD OF SCI

Application of PRTFDC1 gene in promoting porcine skeletal muscle satellite cell differentiation, myotube formation and skeletal muscle growth and development

PendingCN121518581AMicrobiological testing/measurementGenetically modified cellsBiotechnologySkeletal Muscle Satellite Cells
The invention discloses application of a PRTFDC1 gene in promoting porcine skeletal muscle satellite cell differentiation, myotube formation and skeletal muscle development. According to the invention, overexpression and knock-down experiments of the PRTFDC1 are respectively carried out in pig MuSCs, and immunofluorescence, qRT-PCR and Western blot analysis are carried out on cells subjected to induced differentiation, so that the PRTFDC1 is effectively shown to play a positive regulation role in a pig MuSCs differentiation process, and a new marker gene which can be applied to pig molecular breeding and growth performance evaluation is provided; and a novel molecular target and a novel detection tool are provided for muscle development regulation and biological breeding.
Owner:JIANGSU AGRI ANIMAL HUSBANDRY VOCATIONAL COLLEGE

High-affinity PADI2 antibody and preparation method thereof

The invention relates to the field of antibodies, and particularly provides a high-affinity PADI2 antibody and a preparation method thereof. According to the invention, eight high-affinity PADI2 antibodies are obtained by screening through an antibody pair screening experiment, and the antibodies can be specifically combined with PADI2 protein and have high affinity. Experimental results show that the antibody disclosed by the invention shows excellent binding capacity in ELISA (Enzyme-Linked Immunosorbent Assay), Western blot and cell experiments, and can be used for detecting, diagnosing and treating PADI2 related diseases.
Owner:BEIJING GUANGHUI TIANCHENG MEDICAL TECH CO LTD

Monoclonal antibodies to eosinophil peroxidase and uses thereof

The present application relates to the field of biotechnology and in vitro diagnosis, and relates to a monoclonal antibody of eosinophil peroxidase and application thereof. The antibody of the present application is mainly used for detecting the content of eosinophil peroxidase by western blot. The monoclonal antibody of eosinophil peroxidase of the present application is prepared by immunizing Balb / c mice with recombinant eosinophil peroxidase as an immunogen, then fusing the spleen with myeloma cells NS1, and screening and purifying. The prepared monoclonal antibody of eosinophil peroxidase specifically recognizes the linear epitope of eosinophil peroxidase.
Owner:AUTOBIO DIAGNOSTICS CO LTD

Liver cancer cell line with LAPTM5 gene stable overexpression as well as construction method and application of liver cancer cell line

PendingCN120249216AGenetically modified cellsPeptidesHepatoma cell lineLentivirus
The invention discloses a liver cancer cell line with LAPTM5 gene stable overexpression as well as a construction method and application of the liver cancer cell line. According to the invention, a human liver cancer cell line Huh7-LAPTM5 overexpression cell line is successfully constructed through a lentivirus transfection technology. Protein expression is detected through Western blot, and it is proved that overexpression cells are successfully constructed. Meanwhile, experiments prove that under the normal culture condition, LAPTM5 overexpression has no obvious influence on the processes of cell proliferation, apoptosis and the like. The Huh7-LAPTM5 overexpression cell line constructed by the invention plays a biological role by promoting a cell autophagy process, and the cell line is suitable for basic clinical cross correlation researches based on autophagy, drug resistance and the like.
Owner:ZHONGSHAN HOSPITAL FUDAN UNIV

Application of compound fresh bamboo juice in guinea pig asthma model

The invention discloses an application of a compound fresh bamboo juice in a guinea pig asthma model, and relates to the field of medicines, a Hartley albino guinea pig is adopted to construct an OVA induced asthma model, and the model is verified to be successfully constructed through evaluation indexes; evaluating the efficacy of the successfully constructed guinea pig asthma model by using different doses of compound fresh bamboo juice, and evaluating the efficacy of the guinea pig asthma model by scoring guinea pig capsaicin stimulation cough inducing times, alveolar lavage fluid leukocyte number, inflammatory factor content, lung tissue hydroxyproline content and behavioral signs. And evaluating the treatment effect of the compound fresh bamboo juice through Western blot detection, lung histopathology and immunofluorescence staining indexes. In a guinea pig asthma model, high-dose and medium-dose groups of the compound fresh bamboo juice can be used for remarkably reducing the number of cough, reducing the number of white blood cells in alveolar lavage fluid and the level of inflammatory factors and inhibiting pulmonary fibrosis, and a good treatment effect is shown. The results provide powerful support for potential application of the compound fresh bamboo juice in asthma treatment.
Owner:JIANGXI JIUHUA PHARMA +1

Application of 2-indolecarboxylic acid in preparation of tumor targeted therapy drugs

The invention provides an application of 2-indolecarboxylic acid in preparation of a tumor targeted therapy drug. Through computer-aided drug design and molecular docking optimization, 2-indolecarboxylic acid shows high binding energy to an HER2 kinase structural domain. In HER2 positive NCI-N87 gastric cancer cells, the 2-indolecarboxylic acid significantly inhibits cell proliferation and induces G0 / G1 phase arrest and apoptosis. The mechanism research shows that the 2-indoleformic acid can play a role by regulating and controlling the HER2 / Akt / beta-catenin pathway (Western blot verification). In an NCI-N87 cell tumor-bearing nude mouse model, 2-indolecarboxylic acid (15 mg / kg) significantly inhibits tumor growth, and does not cause obvious weight loss or organ toxicity. Immunohistochemical analysis shows that the positive rate of a tumor tissue proliferation marker Ki-67 in a treatment group is remarkably reduced, and apoptosis detection shows that the proportion of TUNEL positive cells is remarkably increased, which indicates that 2-indolecarboxylic acid inhibits tumor cell proliferation and promotes apoptosis at the same time. The invention provides a lead compound with clinical transformation potential for HER2 targeted therapy.
Owner:INST OF MODERN PHYSICS CHINESE ACADEMY OF SCI

Application of berberine in preparation of medicine for resisting porcine reproductive and respiratory syndrome virus

The invention discloses application of berberine in preparation of drugs for resisting porcine reproductive and respiratory syndrome virus, and belongs to the field of antiviral application. The invention relates to an application of berberine in preparation of in-vitro and in-vivo drugs for resisting porcine reproductive and respiratory syndrome virus. According to the invention, berberine is used for pre-incubation treatment of Marc-145 cells susceptible to porcine reproductive and respiratory syndrome virus infection, Western blot and RT-qPCR detection show that berberine can significantly inhibit infection of the porcine reproductive and respiratory syndrome virus on Marc-145 in vitro, and in addition, berberine can also significantly inhibit infection of the porcine reproductive and respiratory syndrome virus on piglets in vivo. The result shows that the berberine has the effect of resisting the porcine reproductive and respiratory syndrome virus, and the berberine can be used for preparing the medicine for resisting the porcine reproductive and respiratory syndrome virus infection so as to prevent the occurrence of the porcine reproductive and respiratory syndrome disease. The invention provides a medicine source for preventing the porcine reproductive and respiratory syndrome, and has important significance for preventing and treating the porcine reproductive and respiratory syndrome virus.
Owner:HEILONGJIANG BAYI AGRICULTURAL UNIVERSITY

High-affinity mouse anti-carp IgM monoclonal antibody, hybridoma cell strain and application thereof

The invention relates to a high-affinity mouse anti-carp IgM (Immunoglobulin M) monoclonal antibody, a hybridoma cell strain and application of the high-affinity mouse anti-carp IgM monoclonal antibody and the hybridoma cell strain. The mouse anti-carp IgM monoclonal antibody is secreted by the hybridoma cell strain CcM-12A12G11C9C7 with the preservation number of CCTCC (China Center For Type Culture Collection) NO: C2024236. The mouse anti-carp IgM monoclonal antibody can be used for Western blot and ELISA specific detection of IgM in a carp secretion form, and can be used for flow cytometry identification of carp IgM + B cells. The monoclonal antibody is good in specificity and high in sensitivity, can be used for structural analysis, immune response level detection and the like of carp IgM, and lays a foundation for in-depth study of a carp immune system and establishment of a vaccine immune effect evaluation method taking the antibody level as an index.
Owner:HUAZHONG AGRI UNIV

Construction method of efficient genetic transformation regeneration system of cabbages

The invention discloses a construction method of a cabbage efficient genetic transformation regeneration system. The method comprises the following steps: (1) culturing aseptic seedlings; (2) preparing an explant; (3) preparing agrobacterium dip dyeing liquid carrying a target carrier; (4) carrying out dip dyeing and co-culture on the explants; (5) callus differentiation and bud induction; and (6) growing seedlings and rooting, and carrying out PCR (Polymerase Chain Reaction) and Western blot identification on the obtained plants. By optimizing key parameters and vector selection of the cabbage genetic transformation method, compared with the prior art, the method has the advantages that antibiotics are not needed for preliminary screening in the regeneration process, hormone is not needed in the rooting process, the cost is low, the transformation period is short (greater than or equal to 45 days), the regeneration frequency is high, the transformation efficiency is high, and subsequent related protein research work can be conveniently carried out; the important value is realized on shortening the cabbage gene function research period.
Owner:GANSU AGRI UNIV

Application of ailanone in preparation of medicine for treating cat infectious peritonitis

PendingCN121622659AOrganic active ingredientsAntiviralsFeline calicivirus infectionWestern blot
The invention discloses application of ailanone in preparation of a medicine for treating feline infectious peritonitis, and belongs to the field of biological medicine. The change of the expression quantity of the feline infectious peritonitis virus N protein is detected through indirect immunofluorescence and western blot, and the change of the quantity titer of the feline infectious peritonitis virus is detected through TCID50. Results show that ailanone has an obvious effect of resisting the feline infectious peritonitis virus, the inhibitory effect on the feline infectious peritonitis virus is enhanced along with increase of the drug concentration, and the half effective concentration is 0.09 mu M. Meanwhile, drug inhibition tests show that ailanone has no inhibition effect on feline calicivirus. The invention provides a novel therapeutic drug and strategy for treating feline infectious peritonitis.
Owner:HUAZHONG AGRI UNIV

Breast cancer cell ferroptosis sensitivity detection method based on down-regulated CDC20

The invention discloses a down-regulated CDC20-based breast cancer cell ferroptosis sensitivity detection method, and belongs to the technical field of detection methods, and the method comprises the following steps: cell grouping and transfection: culturing a breast cancer cell system, and setting an experimental group and a transfection negative control group; wherein in the experimental group, CDC20-siRNA transfection is carried out on breast cancer cells; an experimental group with the CDC20 relative expression quantity lower than 0.5 is screened through Western blot to serve as an optimal experimental group, and subsequent experiments are conducted; and then ferroptosis induction and ferroptosis induction and inhibition are carried out, and related detection is carried out. According to the invention, CDC20 is used as a regulation target of breast cancer ferroptosis sensitivity for the first time, and the target limitation of a traditional method is broken through; and the reliability and repeatability of a detection result are ensured through a standardized operation process and closed-loop verification logic.
Owner:DALIAN MEDICAL UNIVERSITY

Pseudosciaena crocea IRE1 alpha protein polyclonal antibody as well as preparation method and application thereof

The invention relates to the technical field of genetic engineering, in particular to a small yellow croaker IRE1 alpha protein polyclonal antibody as well as a preparation method and application thereof. The invention provides a preparation method of a small yellow croaker IRE1alpha protein polyclonal antibody. The preparation method comprises the following steps: cloning an open reading frame of a small yellow croaker IRE1alpha gene, and constructing a small yellow croaker IRE1alpha gene recombinant expression vector pET-30a (+)-IRE1alpha. Performing inducible expression and purification on the recombinant protein of the Larimichthys polyactis IRE1 alpha, and performing specific analysis on Western Blot; and preparing the polyclonal antibody of the small yellow croaker IRE1 alpha. According to the invention, a recombinant expression vector of the small yellow croaker IRE1alpha gene is constructed by adopting molecular biology and genetic engineering methods, purified recombinant protein is obtained through induced expression and affinity chromatography, the prepared polyclonal antibody can be used for expression detection of the small yellow croaker IRE1alpha protein, and an important material foundation is laid for research on functions and action mechanisms of the small yellow croaker IRE1alpha protein.
Owner:ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES

PROTAC protein targeted degradation agent and application thereof

The invention belongs to the field of pharmacy, and relates to a protein degradation targeting chimera (PROTAC) taking PD-L1 as a target spot and medical application thereof. The protein degradation targeting chimera is a compound composed of a target protein ligand, an E3 ligase ligand and a linker and a pharmaceutically acceptable salt of the compound, the specific structure of the compound is as shown in a formula I, and substituent groups are described in the specification in detail. A Western blot experiment (Western Blot) shows that the compound disclosed by the invention can be used for degrading the PD-L1 protein in a targeted manner.
Owner:SHENYANG PHARMA UNIV

Method for researching effect and mechanism of GOT1 in lung adenocarcinoma cells

The invention discloses an effect and mechanism research method of GOT1 in lung adenocarcinoma cells, and relates to the technical field of biomedical research. Comprising the following steps: S1, cell culture and protein extraction: culturing Bas-2b, A549, H1573 and Calu-3 cells, cleaning the cells with PBS, performing pyrolysis treatment, collecting supernate as a total protein solution, and determining the protein concentration per tube by BCA; s2, carrying out Western Blot experiment, taking 10 micrograms of protein as a loading amount, carrying out electrophoresis, transferring a film, sealing, incubating a primary antibody and a secondary antibody, exposing and developing, and storing a picture; the experiment is repeated for multiple times, and Image J is used for analyzing the strip exposure diagram. According to the method disclosed by the invention, effective knock-down of GOT1 in A549 and H1573 cells is successfully realized through the steps of screening proper cell screening concentration by puromycin, exploring an MOI value, transfecting lentivirus, screening puromycin and the like; the invention provides an important tool and method for follow-up research on the function of GOT1 in lung adenocarcinoma cells.
Owner:THE SECOND AFFILIATED HOSPITAL OF CHONGQING MEDICAL UNIV

Application of lncPRPD regulation and control of GLRX5 in improvement of pork quality

The invention provides application of lncPRPD regulation and control on GLRX5 in improving pork quality, the lncPRPD is lncRNA obtained by performing whole transcriptome sequencing analysis on longissimus dorsi muscle of Laiwu pigs, and the nucleotide sequence of the lncPRPD is as shown in SEQ ID NO: 1. The method comprises the following steps: firstly, synthesizing a small interfering RNA (Ribonucleic Acid) of the lncPRPD / GLRX5 and constructing a gene overexpression vector of the lncPRPD / GLRX5; then, detecting the combination of the lncPRPD and the GLRX5 protein by utilizing a Pulldown experiment and a Western blot experiment; the expression quantity of the GLRX5 is detected after interference and overexpression of the lncPRPD, and it is found that the lncPRPD can promote expression of the GLRX5 protein level; through detection experiments such as qPCR (quantitative polymerase chain reaction), Western blot, oil red O staining and the like, the lncPRPD and the GLRX5 are detected to be capable of promoting the differentiation of the pig precursor fat cells.
Owner:INST OF ANIMAL SCI & VETERINARY MEDICINE SHANDONG ACADEMY OF AGRI SCI