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175 results about "Blot" patented technology

A blot, in molecular biology and genetics, is a method of transferring proteins, DNA or RNA, onto a carrier (for example, a nitrocellulose, polyvinylidene fluoride (PVDF) or nylon membrane). In many instances, this is done after a gel electrophoresis, transferring the molecules from the gel onto the blotting membrane, and other times adding the samples directly onto the membrane. After the blotting, the transferred proteins, DNA or RNA are then visualized by colorant staining (for example, silver staining of proteins), autoradiographic visualization of radioactive labelled molecules (performed before the blot), or specific labelling of some proteins or nucleic acids. The latter is done with antibodies or hybridization probes that bind only to some molecules of the blot and have an enzyme joined to them. After proper washing, this enzymatic activity (and so, the molecules we search in the blot) is visualized by incubation with proper reactive, rendering either a colored deposit on the blot or a chemiluminiscent reaction which is registered by photographic film.

Human lactoferrin indirect competitive enzyme-linked immunosorbent assay kit and application thereof

The invention relates to a human lactoferrin indirect competitive enzyme-linked immunosorbent assay kit and application thereof. The kit comprises an elisa plate, an anti-human lactoferrin antibody, an elisa secondary antibody, a confining liquid, a color developing agent and a stop solution. A nitrocellulose membrane in Western-Blot is replaced by an elisa plate with non-specific adsorption protein, and along with the addition of a sample, an anti-human lactoferrin antibody and an enzyme-labeled secondary antibody, a lactoferrin-primary antibody-secondary antibody compound is formed. And adding a color developing agent to carry out color development reaction, wherein the absorbance value of the sample is positively correlated with the content of the lactoferrin contained in the sample. And calculating the content of human lactoferrin in the sample according to the absorbance value. The method for detecting the human lactoferrin in the fermentation liquor has the advantages of reality, economy, high efficiency and the like.
Owner:JIANGNAN UNIV

Universal visual virus molecular imprinting sensor modified with anti-protein coating

The invention constructs a universal visual virus molecularly imprinted sensor modified with an anti-protein coating, the sensor uses magnetic Fe3O4 as an imprinting carrier, HBV as a template and dopamine as a functional monomer for imprinting to obtain a molecularly imprinted polymer with a polydopamine coating (PDA), and before the template is removed, the molecular imprinting polymer with the polydopamine coating (PDA) is separated from the template, so that the molecular imprinting polymer with the polydopamine coating (PDA) is obtained. The surface of the PDA layer is covered with a slightly cross-linked PMEO2MA layer through a water precipitation polymerization method, and the polymer can effectively reduce non-specific protein adsorption; and finally, exposing an imprinting cavity of the PDA layer by eluting the template molecules. On the other hand, potassium permanganate (KMnO4) is used as a signal output molecule, and when no target object exists, the strong reducibility of PDA can enable KMnO4 entering the imprinting cavity to be subjected to a color fading reaction. Based on the principle, high-specificity colorimetric visual detection on the HBV is realized. The sensor can be combined with a smart phone through naked eyes to carry out visual semi-quantitative analysis and carry out quantitative detection through ultraviolet rays, and the universality of the sensor can be confirmed through a template replacement experiment. And meanwhile, the sensor shows an excellent cost advantage. The diagnosis which is high in sensitivity, low in cost and capable of achieving rapid visualization shows huge potential in practical application, and particularly has important application value in infectious disease screening and monitoring work in areas with limited resources.
Owner:XIANGTAN UNIV

Porcine reproductive and respiratory syndrome virus dimer nucleocapsid protein and application thereof

The invention relates to a porcine reproductive and respiratory syndrome virus (PRRSV) dimer nucleocapsid (N) protein (PRRSV N-dimer) and application thereof. The PRRSV N-dimer is formed by connecting N protein amino acid sequences in series through (GGGGS) 3linker. According to the invention, a GGGGS 3linker nucleotide sequence is connected in series with a PRRSV NADC30-like strain N protein gene sequence to construct a prokaryotic expression vector, and a recombinant PRRSV N-dimer is expressed in escherichia coli. Through SDS-PAGE (Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis) and Western-blot detection, the recombinant PRRSV N-dimer obtains soluble expression and can be subjected to a specific reaction with PRRSV positive serum; through detection by a freeze thawing method and an ELISA (Enzyme-Linked Immunosorbent Assay) method, compared with a PRRSV monomer N protein (PRRSV N-monomer), the PRRSV N-dimer protein can be stably preserved, and the immunoreactivity is better. A PRRSV antibody indirect enzyme-linked immunosorbent assay (iELISA) detection method is established on the basis of a PRRSV N-dimer, and can be applied to diagnosis and monitoring of PRRSV infection.
Owner:HENAN ACAD OF AGRI SCI

Monoclonal antibody against soluble transferrin receptor as well as preparation method and application thereof

The invention discloses an anti-soluble transferrin receptor monoclonal antibody as well as a preparation method and application thereof, and belongs to the technical field of antibody detection. According to the antibody, an L101-F760 amino acid sequence of human TfR protein is used as an immunogen, specific screening of B cells, reverse RNA (Ribonucleic Acid) into cDNA (Complementary Deoxyribose Nucleic Acid) through RT-PCR (Reverse Transcription-Polymerase Chain Reaction), then homologous recombination is performed to construct heavy and light chain vectors, and finally multiple rounds of screening are performed to obtain the purified sTfR monoclonal antibody, so that the sTfR protein can be specifically recognized, and the antibody has the advantages of high specificity and high sensitivity and can be widely applied to the field of immunotherapy. The natural sTfR protein in human serum can be recognized. The kit is applied to the detection and screening fields of immunohistochemistry, immunofluorescence chemistry, western blot, co-immunoprecipitation, indirect ELISA, flow cytometry and the like, especially has low background and no interference of non-specific binding impure protein during immunohistochemistry detection, can obtain more accurate detection and evaluation results, avoids interference of false positive and false negative, and has high detection accuracy. The method has great application value in the aspects of basic scientific research and clinical pathology detection.
Owner:HANGZHOU STAR BIOTECHNOLOGY CO LTD

Immunoblotting image recognition method and system based on deep learning

The invention discloses an immunoblotting image recognition method and system based on deep learning, relates to the field of bioinformatics and computer vision, and solves the problem that it is difficult to extract various related features of protein from an image through an image recognition technology. The method solves the technical problem that a prediction value obtained by predicting a protein expression level obtained by a deep learning model through various protein feature data is difficult to correct. The method comprises the following steps: capturing a strip image on a film by using a high-resolution scanner; recognizing the preprocessed image through an image recognition technology, and extracting features related to the immunoblotting; predicting the feature data through a correspondingly trained deep learning model to obtain an area prediction value, a gray scale difference degree prediction value and a relative brightness prediction value; corresponding weights are given to the three prediction correction values for addition, and a comprehensive prediction correction value is obtained; and comparing the comprehensive expression evaluation value with a preset threshold value to judge the protein expression level.
Owner:HUNAN ZHONGRUI MUTUAL TRUST MEDICAL TECH CO LTD

Disease marker subtype classification determination method

PendingCN120279987ABiostatisticsBiological modelsImmunoblot AssayAlgorithm
The invention provides a disease marker subtype classification determination method, and belongs to the technical field of disease marker subtype classification, and the method comprises the following steps: firstly, extracting protein from a patient sample, separating through a dimensional electrophoresis technology, and obtaining a protein expression matrix through Coomassie brilliant blue dyeing and an image analysis system; differentially expressed proteins are determined, and sequence information is obtained through mass spectrum identification. And extracting network node features by using a graph neural network method, and carrying out function annotation analysis and path identification. Patient samples are grouped by using a multi-level clustering method, and feature extraction and optimization are realized by training an embedded neural network model. And inputting the extracted features into a pre-trained function enrichment analysis model, and determining the molecular pathway features of the disease subtype. And finally, the reliability of a classification result is confirmed through an immunoblotting technology and independent patient queue verification, subtype classification of the disease markers is completed, and the problem that in the prior art, the recognition degree of subtype classification of the disease markers is not high enough is solved.
Owner:QINGDAO RAISECARE BIOTECHNOLOGY CO LTD

Screening method of sicca syndrome markers based on conjunctival blotting

The invention discloses a screening method of sicca syndrome markers based on conjunctival blotting, and relates to the technical field of biomedicine, and the screening method comprises the following steps: S1, conjunctival noninvasive sampling; s2, RNA (Ribonucleic Acid) sequencing and pathway screening; s3, machine learning modeling is carried out; s4, clinical verification. According to the invention, biomarker screening is realized, and similar biomarker screening can also be realized through tear sample analysis or other ophthalmic samples; however, the alternative schemes have the defects of relatively strong invasiveness, complex operation and the like; sampling is carried out through the conjunctival imprinting technology, invasive operation such as biopsy and blood drawing is avoided, and pain and discomfort are reduced; differential expression genes can be accurately identified, and the sensitivity and the accuracy are relatively high; through immune cell infiltration analysis, the relationship between the immune cell subpopulation and the sicca syndrome related gene is disclosed, a new perspective is provided for a disease mechanism, and a theoretical basis is provided for a future immune regulation strategy.
Owner:THE FIRST AFFILIATED HOSPITAL OF CHONGQING MEDICAL 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

Self-cleaning flexible SERS (Surface Enhanced Raman Scattering) blotting membrane for selectively detecting trace microcystin

The invention provides a preparation method and application of a high-sensitivity imprinted membrane for selectively detecting microcystic toxins (MC-LR) in water based on surface enhanced Raman scattering. The preparation method comprises the following steps: step 1, preparation of silver nanocubes; step 2, preparation of Ag NCs-coated TiO2 nanoparticles; step 3, preparation of an Ag NCs-coated TiO2 / PDMS composite membrane; according to the invention, a membrane technology, an SERS technology and MIT are combined, so that the prepared product has excellent physical and chemical properties of a PDMS membrane, high sensitivity of an SERS detection technology and high selectivity of MIT; the invention provides a new method for SERS detection and widens the application range.
Owner:HAINAN NORMAL UNIV

Preparation method of high-activity oyster-derived small molecule peptide

PendingCN121320478APeptide preparation methodsFermentationNeutral proteaseCrassostrea rivularis
The invention relates to the field of biological product preparation, in particular to a preparation method of high-activity oyster-derived small molecule peptide. The method comprises the following steps: selecting fresh meat of pacific oysters or ostrea rivularis as a raw material, degreasing, adding water, crushing, filtering, sequentially carrying out compound protease and neutral protease hydrolysis, implanting a sensor and a probe in hydrolysis, monitoring in real time, and dynamically adjusting reaction conditions; adding a targeting microcapsule wrapping an ascorbic acid glutathione composite protective agent, and releasing the protective agent when a target peptide signal is detected; separating and concentrating through a gradient membrane, and purifying the target peptide by using a molecular imprinting chromatographic column; carrying out alkaline hydrolysis on enzymatic hydrolysis residues to prepare chitosan for cyclic utilization, carrying out enzymatic hydrolysis on residual residues to generate collagen peptide, treating and recycling wastewater, and recovering protease. According to the method, the technical effects that the high-activity and high-purity target peptide is prepared, the resource utilization rate is increased, the production cost is reduced, the target peptide has good oxidation resistance and biological activity, the intestinal enzyme degradation rate is reduced after glycosylation modification, and the blood half-life period is prolonged are achieved.
Owner:HEBEI JINMU PHARM GRP CO LTD

Preparation method of magnetic molecular imprinting-aptamer sandwich fluorescence sensor for detecting kanamycin

The invention designs a preparation method of a magnetic molecular imprinting-aptamer sandwich fluorescence sensor for detecting kanamycin. And the magnetic molecular imprinting composite material Fe3O4 (at) UiO-66 (at) MIP with a specific adsorption target object KANA is prepared. When a detection target object KANA exists, the Fe3O4-coated UiO-66-coated MIP can rapidly and specifically capture the KANA in a short time. Then, a DNA single-stranded aptamer marked with FAM is added, after the aptamer and KANA captured on the surface of the Fe3O4 at-UiO-66 at-MIP are subjected to specific recognition and magnetic separation again, an aptamer probe can enter sediment along with the magnetic composite material Fe3O4 at-UiO-66 at-MIP, the fluorescence intensity of supernate is reduced, and quantitative analysis is conducted according to the fluorescence difference value delta F. When the KANA does not exist, the aptamer cannot enter the precipitate, so that the fluorescence signal intensity is not changed. Double recognition is formed through specific binding of an MIP imprinting cavity and the aptamer, matrix interference is reduced by means of fluorescence characteristics and magnetic separation capacity, the method has the advantages of being high in adsorption capacity, specific in recognition, high in sensitivity, wide in detection range, good in selectivity and the like, and rapid and accurate detection of KANA in a complex sample can be achieved.
Owner:HENAN UNIVERSITY OF TECHNOLOGY

Fluorescence coding probe and single cell immunoblotting method

The invention discloses a fluorescence coding probe and a single cell immunoblotting method. The nucleic acid tag comprises oligonucleotide chains f1, f2... fn, n is greater than 2, each oligonucleotide chain contains a fluorophore and a quenching group, partial sequences of the next oligonucleotide chain and the previous oligonucleotide chain are complementarily paired, and the quenching group of the next oligonucleotide chain quenches the fluorophore of the previous oligonucleotide chain. Also disclosed is a quenching tag comprising a complementary strand capable of complementary pairing with an oligonucleotide strand in a nucleic acid tag. The invention discloses a fluorescent coding probe comprising a nucleic acid label, a preparation method of the fluorescent coding probe and application of the fluorescent coding probe in a multi-target single-cell immunoblotting technology, an immunofluorescence technology and an immunohistochemical staining technology. According to the fluorescence-coded multi-target single-cell immunoblotting method disclosed by the invention, multi-target protein detection can be carried out by utilizing limited fluorescence channels, the signal loss is small, and the expansibility is high.
Owner:SHANGHAI JIAO TONG UNIVERSITY INNER MONGOLIA RESEARCH INSTITUTE

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

African swine fever virus pnp868r protein monoclonal antibody and application thereof

The application belongs to the technical field of biology and particularly relates to an African swine fever virus pNP868R protein monoclonal antibody and a preparation method and application thereof. The application takes the pNP868R protein prepared by eukaryotic expression as an antigen, immunizes BALB / c mice, uses hybridoma technology, and obtains a hybridoma cell line capable of stably secreting the anti-African swine fever virus pNP868R protein monoclonal antibody through cell fusion, indirect ELISA method screening, limited dilution method subcloning and hybridoma continuous passage culture. The application also obtains an African swine fever virus pNP868R protein monoclonal antibody. The monoclonal antibody has high affinity with the pNP868R protein and can be applied in Western-Blot experiments, indirect immunofluorescence experiments, immunoprecipitation experiments and the capture of proteins interacting with pNP868R, and can be used for the diagnosis, prevention or treatment of African swine fever.
Owner:LANZHOU UNIV

Preparation method and application of molecularly imprinted composite filler

The invention relates to a preparation method of a molecularly imprinted composite filler, and relates to the technical field of analytical chemistry and environmental monitoring technologies, the preparation method of the molecularly imprinted composite filler comprises the following steps: firstly dissolving template molecules and a first monomer, then adding a cross-linking agent and an initiator, carrying out deoxidation and water bath, and then centrifuging to obtain a template molecule solution; and eluting the precipitate with a flushing fluid, and drying to obtain the microspheres. And dissolving the ionic liquid, uniformly stirring with the obtained microspheres, and carrying out centrifugal separation, washing and vacuum drying to finally prepare the molecularly imprinted composite filler. Finally, the hydrophobic property of the molecularly imprinted composite filler is improved through modification of the ionic liquid and the second monomer. The method provided by the invention has excellent hydrophobic effect and enrichment capability on a target object.
Owner:SHENZHEN TENGBIAO TESTING CO LTD

A trimodal detection method for ethyl carbamate based on molecular imprinting regulation of Co@MOF-MIP enzyme activity switch

A trimodal detection method for ethyl carbamate based on molecular imprinting regulation of Co@MOF-MIP enzyme activity switch, comprising the following steps: Step 1: Determine the absorbance value A of the mixed reaction solution of ethyl carbamate at 650nm n Step 2: Under the excitation wavelength of 365nm, the fluorescence intensity value F of the mixed reaction solution of ethyl carbamate at 440nm is measured. n Step 3: Determine the temperature T of the mixed reaction solution of ethyl carbamate under 660nm laser irradiation n :Step 4: Construct a linear regression equation for the concentration of ethyl carbamate; Step 5: Take the sample to be tested and repeat steps 1, 2 and 3 to obtain the absorbance signal of the colorimetric mode, the fluorescence intensity signal of the fluorescence mode and the temperature signal T of the photothermal mode n The synthesis process of Co@MOF‑MIP nanozyme is relatively simple, the raw materials are readily available and low-cost, and it does not require complex processes and expensive equipment. It has high stability and high peroxidase activity and fluorescence properties, which meet the needs of subsequent multimodal detection of EC.
Owner:INTELLIGENT MFG INST OF HFUT +1

Full-automatic micro-protein immunoblotting analyzer and analysis method thereof

The invention relates to the technical field of western blot analysis, and discloses a full-automatic micro western blot analyzer and an analysis method thereof. The full-automatic micro protein immunoblotting analyzer comprises a full-automatic control system, a horizontal control system, a protein separation system, a temperature control system, a liquid path control system, an ultraviolet curing module, an antibody incubation module and a chip scanning module, the horizontal control system controls the protein separation system to horizontally move and correct; the protein separation system carries out protein separation; the temperature control system is used for carrying out constant-temperature control and temperature adjustment; the liquid path control system is used for filling a constant-temperature reagent into the protein separation system and extracting waste liquid; the ultraviolet light curing module is used for carrying out in-situ immobilization on the separated protein; the antibody incubation module is used for performing antibody incubation on the fixed protein marker; and the chip scanning module is used for chip result reading and data processing. According to the invention, full-automatic control, accurate temperature control and automatic horizontal correction can be realized.
Owner:SHANGHAI JIA DOCTOR TECHNOLOGY CO LTD

A peanut allergen polypeptide, its screening method and application

This invention provides a peanut allergen peptide, its screening method, and its application. Phage display technology is used to screen for conformational epitopes of Ara h5 and Ara h8 proteins. Polyclonal IgE and IgG antibodies against the target proteins are prepared by immunizing BALB / c mice with Ara h5 and Ara h8 proteins as antigens. ELISA and dotted immunoblotting detection revealed that the titers of polyclonal IgG and IgE against Ara h5 and Ara h8 proteins were 1:64000 and 1:320, respectively, and the antibodies exhibited specificity. Using the antibodies as targets, phage display technology screened the conformational epitope sequences of Ara h5 protein as WETIYSR and FHWWYLK. The conformational epitopes of Ara h8 protein were FPYMKFV, FPYMKFR, SMFARID, and SFHWWLF.
Owner:CHINA AGRI UNIV

Molecularly imprinted electrochemical-photoelectrochemical dual-mode sensor for detecting tetracycline as well as preparation method and application of molecularly imprinted electrochemical-photoelectrochemical dual-mode sensor

The invention provides a molecularly imprinted electrochemical-photoelectrochemical dual-mode sensor for detecting tetracycline as well as a preparation method and application of the molecularly imprinted electrochemical-photoelectrochemical dual-mode sensor, and relates to the technical field of detection. According to the molecularly imprinted electrochemical-photoelectrochemical dual-mode sensor for detecting tetracycline, provided by the invention, the tetracycline is accurately detected by utilizing a quenching effect of a steric hindrance effect caused by specific binding of tetracycline and an imprinted cavity on electrochemical and photoelectric signals, and the molecularly imprinted electrochemical-photoelectrochemical dual-mode sensor has the advantages of wide detection range, high sensitivity and high accuracy. Wherein the electrochemical detection linear range is 5.010 <-14 > to 1.010 <-9 > mol / L, and the detection limit is 2.110 to 14 mol / L; the linear range of photoelectrochemical detection is 5.010 <-12 >-5.010 <-6 > mol / L, the detection limit is 2.710 <-13 > mol / L, and the method is successfully used for detecting the content of tetracycline in water around livestock and poultry farms and has good analysis performance.
Owner:JIANGSU UNIV

SARS-COV-2 immunogenic compositions

Disclosed herein are compositions comprising protein antigens and RNA encoding the same (eg., compositions comprising protein antigens and RNA encoding antigens) that can be used to induce an immune response against SARS-CoV-2. Also disclosed herein are immunogenic compositions and medical preparations comprising the same, and methods of making and using the same. In some embodiments, the technologies provided herein can be used to address and / or overcome immune imprinting in SARS-CoV-2.
Owner:BIONTECH SE +1

Anti-mouse IgG nano antibody and alkaline phosphatase fusion protein as well as preparation method and application thereof

PendingCN120535657ABacteriaAntibody mimetics/scaffoldsEscherichia coliChemical labeling
The invention relates to an anti-mouse IgG nano antibody and alkaline phosphatase fusion protein as well as a preparation method and application thereof. The anti-mouse IgG nano antibody and alkaline phosphatase fusion protein sequentially comprises an anti-mouse IgG nano antibody, a connecting peptide and shewanella alkaline phosphatase from an amino terminal to a carboxyl terminal. The fusion expression of the anti-mouse IgG nano antibody and shewanella alkaline phosphatase in escherichia coli is designed, so that various defects of a chemical labeling method can be avoided, a high-purity and high-specific activity enzyme-labeled antibody can be prepared, the preparation period of a second antibody can be shortened, the preparation process of the second antibody can be simplified, the stability of batches can be improved, and the dependence on experimental animals can be reduced. The prepared anti-mouse IgG nano antibody and alkaline phosphatase fusion protein can be applied to the field of immunodetection, such as a chemiluminescence immunoassay method, an enzyme-linked immunosorbent assay method, an enzymatic fluorescence immunoassay method, an immunoblotting method and the like.
Owner:GENE TECH SHANGHAI COMPANY

An anti-MSLN antibody and its application

This invention discloses an anti-MSLN antibody and its applications. The anti-MSLN antibody comprises a heavy chain variable region and a light chain variable region. The heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3. The amino acid sequences of HCDR1, HCDR2, and HCDR3 are shown in SEQ ID NO:7, 8, and 9, respectively; and the amino acid sequences of LCDR1, LCDR2, and LCDR3 are shown in SEQ ID NO:4, 5, and 6, respectively. The antibody provided by this invention has good affinity and specificity for MSLN antigens, effectively detecting target proteins in cells overexpressing MSLN. It can also be widely used as a core ingredient in the development of high-sensitivity MSLN detection kits such as ELISA, Western blotting, and immunohistochemistry.
Owner:SHANGHAI TECH UNIV +1

Efficient ribosome imprinting sequencing library construction method

The invention discloses an efficient ribosome imprinting sequencing library construction method, and belongs to the technical field of biology. In the enzyme digestion treatment step, a mixed enzyme system of RNase I and micrococcus nuclease is adopted to treat the cell lysis buffer, and the trinucleotide periodicity of RPFs can be remarkably improved to 80% or above. In the target fragment recovery step, the sucrose gradient ultracentrifugation step is omitted, the RPFs are directly recovered by gel electrophoresis, a plurality of enzyme reaction steps in library construction are optimized and integrated, and streptavidin magnetic beads are used for solid phase purification, so that the whole library construction period is greatly shortened from 7 days to 2.5 days. The method does not depend on special equipment such as ultracentrifugation, and can be completed in a conventional molecular biology laboratory. Besides, a multi-enzyme synergistic RNA terminal one-step modification system is innovatively constructed, single-tube one-step completion is realized, the terminal treatment efficiency and consistency are improved, and a key guarantee is provided for obtaining a high-quality and high-complexity Ribo-seq library.
Owner:YUNNAN UNIV

Fish creatine kinase allergenicity and immune cross reactivity analysis method

The invention provides a fish creatine kinase sensitization and immune cross reactivity analysis method which comprises the following steps: preparing a fish muscle water-soluble protein crude extract, separating 40-45kDa protein components in the fish muscle crude extract through SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis), and screening a target sensitization protein combined with IgE / IgG through immunoblotting; the method comprises the following steps: extracting total RNA of grass carp and synthesizing cDNA, designing a creatine kinase specific primer for amplification to obtain a creatine kinase gene sequence, and carrying out bioinformatics analysis after sequencing verification; the method comprises the following steps: cloning a creatine kinase gene to an expression vector, transforming escherichia coli BL21, performing IPTG induced expression, and performing urea gradient dialysis renaturation on an inclusion body to obtain the recombinant grass carp creatine kinase. Detecting the IgE binding activity of the recombinant grass carp creatine kinase and the serum of the allergic patient through dot hybridization; the IgG cross reactivity of the recombinant grass carp creatine kinase and the blue crab arginine kinase is verified through indirect ELISA; the IgE cross reaction degree of the recombined grass carp creatine kinase and the blue crab arginine kinase is quantified through inhibitory dot hybridization.
Owner:XIAMEN HUAXIA UNIV

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

A self-activation fenton-like composite repair material and a preparation method and application thereof

The application relates to a Fenton-like composite material, in particular to a self-activated Fenton-like composite repair material and a preparation method and application thereof, the composite repair material is CaO2@Fe3O4@BIP-BC, a protein imprinting biochar is used as a matrix, magnetite serving as an activator and calcium peroxide serving as a slow-release oxidant are sequentially loaded on the surface. Compared with the prior art, the application has the advantages that (1) the biochar surface active groups and calcium peroxide improve the high efficiency and long-acting property of the Fenton-like process, (2) the protein imprinting improves the selectivity of the removal of target harmful bacteria and harmful bacteria genes, (3) the biochar simultaneously loads the calcium peroxide and the magnetite, the simultaneous addition of the activator and the oxidant is realized, and the use cost and the operation complexity are reduced. The method solves the problems that the traditional Fenton-like oxidation treatment has poor specific removal effect on harmful bacteria and harmful bacteria genes in the environment, the treatment time is short, the required dosage is high, secondary pollution is serious, and the activator and the oxidant need to be used separately.
Owner:SHANDONG NONGHUAN GREEN PRODUCTS ENVIRONMENTAL TECHNOLOGY CO LTD +1

Method for high-throughput identification of lncrna-encoded peptides and application thereof

ActiveCN115985397BVerify feasibilitySequence analysisInstrumentsProteomics methodsBase J
The application relates to a high-throughput identification method of lncRNA coding peptides and application thereof, and belongs to the technical field of biology. The method combines a translationomics technology and a protein mass spectrometry technology, takes ribosome footprint sequencing data as a starting point, determines an open reading frame located on an lncRNA gene according to a three-base periodicity rule of a sequencing fragment, further constructs a proteogenomics index after systematic filtration, uses a computational proteomics method to detect an lncRNA translation event, and systematically identifies lncRNA coding peptides on the basis. The method has the characteristics of modularity and strong expandability, can be widely used for various types of proteome data, and can high-throughput identify and quantitatively detect lncRNA coding peptides. The application provides a prediction of molecular typing and prognosis of cancer patients based on an lncRNA coding peptide network model, and provides an important reference basis for precise medication of clinical patients.
Owner:SHENYANG PHARMA UNIV

Visual bifunctional subtype influenza virus molecular imprinting sensor as well as preparation method and application thereof

The invention discloses a visual bifunctional subtype influenza virus molecularly imprinted sensor as well as a preparation method and application thereof. The sensor comprises a fluorescent signal core and a molecularly imprinted polymer layer coated outside the fluorescent signal core, the fluorescent signal core is SiO2 microspheres coated with CdTe quantum dots, the molecularly imprinted polymer layer is internally provided with an imprinting cavity capable of specifically recognizing the subtype influenza virus, the three-dimensional space shape of the imprinting cavity is complementary to that of the subtype influenza virus, and the molecularly imprinted polymer layer is coated with CdTe quantum dots. Sialic acid molecules are covalently modified on the inner surface. The sensor can be used for quantitative detection or visual semi-quantitative detection of subtype influenza viruses such as H7N9 and H5N1, and integrates double functions of rapid screening and accurate quantification. The sensor disclosed by the invention is good in preparation reproducibility, high in selectivity, strong in anti-interference capability and suitable for analysis of complex biological samples, and a novel user-friendly and efficient method is provided for on-site monitoring and early warning of influenza viruses.
Owner:YUNNAN NORMAL UNIV

A molecular imprinting electrochemiluminescence sensor, a preparation method thereof and application of the sensor in selective detection of trilobatin

The application belongs to the technical field of electrochemical analysis and detection, and discloses a molecular imprinting electrochemiluminescence sensor, a preparation method thereof and application of the sensor in selective detection of trilobatin. The operation comprises the following steps: (1) a prepared g-C3N4@NiMOF composite material is modified on the surface of a glassy carbon electrode; (2) the g-C3N4@NiMOF / GCE is placed in a phosphate buffer solution containing o-phenylenediamine and TRI, and o-PD and TRI are electropolymerized on the electrode surface through cyclic voltammetry in a three-electrode system to obtain MIP-p(o-PD)-TRI / g-C3N4@NiMOF / GCE; and (3) TRI in the polymerization film is eluted to form a specific imprinting cavity, and a MIP-p(o-PD) / g-C3N4@NiMOF / GCE sensor is obtained. The molecular imprinting electrochemiluminescence sensor has the advantages of simple operation, fast analysis speed, good selectivity and high sensitivity in the detection of TRI.
Owner:CHANGZHOU UNIV

Protein surface oriented imprinting microsphere as well as preparation method and application thereof

The invention discloses a protein surface oriented imprinting microsphere as well as a preparation method and application thereof, and belongs to the technical field of western blot materials. According to the method provided by the invention, the protein surface imprinting material is successfully prepared by virtue of surface directional imprinting and water-phase polymerization, the stability is high, the interaction force with the protein to be detected is strong, and the target protein can be stably and specifically recognized; according to the protein surface oriented imprinting core-shell microsphere prepared by the method provided by the invention, a base sphere is of a dendritic structure, the diameter is 80nm, the surface has abundant wrinkles and high porosity, a shell layer is a polyion liquid macromonomer polymer layer, target protein is oriented and anchored, and the microsphere has abundant recognition sites, excellent dispersity and high particle size uniformity; the provided preparation method of the protein surface oriented imprinting microspheres is simple and easy to operate, the condition is mild, the used reagent is low in toxicity or non-toxic, no toxic or harmful substance is generated, and the preparation method is environment-friendly.
Owner:XIAN UNIV OF SCI & TECH