22 results about "Protein blotting" patented technology

The invention discloses preparation and application of grouper enterospora sporowall protein SWP26 and a polyclonal antibody of the grouper enterospora sporowall protein SWP26, and belongs to the field of animal quarantine, the grouper enterospora sporowall protein SWP26 is obtained by amplifying grouper enterospora genes by using a PCR (Polymerase Chain Reaction) method, shearing a target band and transferring the target band into a pET-32a vector to construct a recombinant plasmid, and then transforming escherichia coli BL21 (DE3) for induced expression. A Ni-NTA affinity chromatography method is used for protein purification, animal immunization and polyclonal antibody purification are carried out on the purified SWP26 recombinant protein, western blot detection is carried out on the purified antibody, an obvious signal appears at about 43kDa, and it is proved that the anti-SWP26 polyclonal antibody can be subjected to a specific reaction with the SWP26 purified protein. The invention clones and identifies the high-abundance spore wall protein SWP26 positioned on the surface of the enterosporidium of the grouper for the first time, belongs to a specific protein of the enterosporidium of the grouper, and can be used as a drug target for treating enterocytozoonosis of the grouper.

The present invention provides a multiomics approach, which integrate single-cell RNA-sequencing (scRNA-seq) and spatiotemporal transcriptomics (ST) offering potential for dissecting transcriptional networks and revealing cell-cell interactions involved in biomolecular pathomechanisms. The present invention also provides a multimodal approach combining high-throughput scRNA-seq and ST to elucidate XLRS-specific transcriptomic signatures in two XLRS-like models with retinal splitting phenotypes, including genetically engineered (Rs1emR209C) mice and patient-derived retinal organoids harboring the same patient-specific p.R209C mutation. Through multiomics transcriptomic analysis, the endoplasmic reticulum (ER) stress / eIF2 signaling, mTOR pathway, and the regulation of eIF4 and p70S6K pathways as chronically enriched and highly conserved disease pathways between two XLRS-like models are identified. Western blots and proteomics analysis validated the occurrence of unfolded protein responses, chronic eIF2α signaling activation, and chronic ER stress-induced apoptosis. Furthermore, therapeutic targeting of the chronic ER stress / eIF2α pathway activation synergistically enhanced the efficacy of AAV mediated RS1 gene delivery, ultimately improving bipolar cell integrity, postsynaptic transmission, disorganized retinal architecture and electrophysiological responses. Collectively, the complex transcriptomic signatures obtained from Rs1emR209C mice and patient-derived retinal organoids using the multiomics approach provide opportunities to unravel potential therapeutic targets for incurable retinal diseases, such as XLRS.

The invention relates to the field of biotechnology and materials, in particular to an improved western blot transfer printing film as well as a preparation method and application thereof. Specifically, the invention relates to a chemically modified polyvinylidene fluoride membrane, and the surface of the membrane is modified with a group as shown in a formula I which is described in the specification. After the protein is transferred to the chemically modified polyvinylidene fluoride membrane, 365 nm ultraviolet rays can be used for irradiation, so that the protein is covalently cross-linked to the membrane. The invention also relates to a preparation method of the chemically modified polyvinylidene fluoride membrane and application of the chemically modified polyvinylidene fluoride membrane in western blot.

This invention discloses a protein blotting device, belonging to the technical field of immunoblotting equipment. It includes: a base, an incubation tray placement section on the base; an incubation tray placed on the incubation tray placement section, with multiple incubation cells spaced apart on the incubation tray; a liquid addition device for respectively introducing detection solutions into the incubation cells and mounted on the base; a liquid aspiration device for respectively aspirating the detection solutions from the incubation cells and mounted on the base; and a blowing and agitating device for blowing air into the incubation cells to cause the detection solutions in the incubation cells to flow relative to the membrane strip placed in the incubation cells. The blowing and agitating device is mounted on the base and has an air outlet structure, which is located above the incubation tray placement section, with the air outlet of the air outlet structure facing the incubation cells of the incubation tray horizontally placed on the incubation tray placement section. The protein blotting device of this invention helps ensure sufficient contact and reaction between the membrane strip and the detection solution while preventing spillage of the detection solution.

The invention relates to the technical field of western blot membranes, in particular to a high-strength western blot membrane based on nanofiber network interpenetration and a preparation method thereof.The high-strength western blot membrane comprises the following raw materials: a polyvinylidene fluoride membrane matrix, a nanofiber skeleton, a pore-foaming agent, an N, N-dimethylacetamide cosolvent and coagulating bath; the preparation method comprises the following steps: selecting a cellulose nanofibril aqueous dispersion with the concentration of 2wt%, carrying out hydrophobic modification on the cellulose nanofibril aqueous dispersion through octyl triethoxy silane, and taking the modified cellulose nanofibril aqueous dispersion as a nanofiber skeleton; the pore-foaming agent is a compound mixed solution of polyvinylpyrrolidone and PEG-400 (Polyethylene Glycol). The cellulose nanofibrils are subjected to surface hydrophobization modification, and a compound pore-foaming agent system is introduced, so that the mechanical property and the porous structure of the membrane are considered, and the good porosity of the membrane is maintained while the overall strength of the membrane is enhanced.

Systems, methods, and components for automated processing and analyzing a biological sample are disclosed herein. Such components include pre-filled gel cards, gel card cassettes and blotting cartridges for use in automated electrophoresis and protein transfer. Systems includes automated electrophoresis and transfer modules that automatically separate proteins from the biological sample into a plurality of bands and transfer the plurality of bands to a membrane using the gel card cassettes and blotting cartridges herein. Compared to conventional methods, the disclosed system and method provide improvements in reduced experiment time, reduced error, improved precision, and reduced consumption of consumables and chemicals.

The invention discloses application of a reagent for detecting LSH in preparation of a chronic obstructive pulmonary disease diagnosis preparation and application of a reagent for overexpressing the LSH in preparation of a medicine for treating the chronic obstructive pulmonary disease. Protein expression of LSH in a patient suffering from the chronic obstructive pulmonary disease is found to be lower than that of healthy people and expression of pure smokers through western blot and RT-qPCR detection for the first time. Therefore, the possibility of using the LSH as the detection marker of the chronic obstructive pulmonary disease is provided. The LSH overexpression adeno-associated virus improves obstructive ventilation dysfunction and large and small airway dysfunction of a model mouse, relieves the emphysema degree, reduces the airway inflammation level and inhibits small airway remodeling, and further analysis on inflammatory cells in the lung shows that the overexpression of the LSH overexpression adeno-associated virus can reduce the proportion of pulmonary interstitial macrophages. The invention provides a new way with a prospect for treating the chronic obstructive pulmonary disease.

The invention provides a polyvinylidene fluoride membrane as well as a preparation method and application thereof, and relates to the technical field of materials. The preparation method of the polyvinylidene fluoride membrane provided by the invention comprises the following steps: coating a membrane casting solution on a base material to form a liquid membrane, staying a carrier in an environment of 30-80 DEG C for 1-5 seconds, immersing the carrier into a coagulating bath, curing to form a membrane, and washing with water to obtain the polyvinylidene fluoride membrane. The preparation method is environment-friendly, simple in process and low in cost, and the prepared polyvinylidene fluoride membrane has a dendritic-spherocrystal alternate porous structure and controllable membrane aperture, and can be used for biological application technologies such as western blot, sterilization and filtration.

The present invention relates to the field of biotechnology and materials, in particular to an improved Western blotting membrane, its method of preparation and use. Specifically, the present invention relates to a chemically modified polyvinylidene fluoride membrane, said membrane being surface modified with a group according to Formula I: Formula I. After transfer of a protein to the chemically modified polyvinylidene fluoride membrane of the present invention, 365 nm UV light can be used to covalently crosslink the protein to the membrane. The present invention also relates to a method of preparation of the chemically modified polyvinylidene fluoride membrane, and its use in Western blotting.

This invention belongs to the field of protein antibody incubation technology and provides a protein blotting processing device, comprising: a base; a limiting post fixedly disposed on the base, one end of the limiting post having a cam component, the cam component having at least one protrusion and / or one concave portion; a turntable rotatably disposed relative to the limiting post, comprising a first fixed plate and a second fixed plate, the cam component being located between the first fixed plate and the second fixed plate, and the cam component and the second fixed plate having a preset distance; at least one first push rod movably disposed on the lower side of the second fixed plate, one end of which can abut against the cam component, the cam component being able to drive the first push rod to reciprocate along its own axis; a reaction tank movably disposed on the upper side of the second fixed plate, and passing through the second fixed plate and movably connected to the other end of the first push rod. By adopting the above structure, the reaction tank can be oscillated simultaneously with the rotation of the turntable, aided by the cam component and the first push rod.

The utility model provides a western blot instrument reagent bottle fixing device. Relates to the technical field of experimental instruments, the western blot instrument reagent bottle fixing device comprises a bottom plate, four vertical plates are fixedly installed at the top of the bottom plate, a same placing plate is fixedly installed at the tops of the four vertical plates, a plurality of reagent bottles are slidably installed on the placing plate, the reagent bottles are in sliding contact with the bottom plate, and the bottom plate is provided with a plurality of reagent bottles. Two U-shaped rods are fixedly mounted at the bottom of the placing plate, two fixing blocks are fixedly mounted at the bottom of the placing plate, a same transfer rod is rotatably mounted between the two fixing blocks, six moving blocks are arranged on the transfer rod in a threaded and sleeving manner, and mounting strips are fixedly mounted on the two sides of the six moving blocks. The reagent bottle fixing device has the advantages that the operation is convenient and fast, a plurality of reagent bottles can be more stably and quickly fixed, and the possibility that the reagent bottles fall off is greatly reduced.

This invention provides a polyvinylidene fluoride (PVDF) membrane, its preparation method, and its applications, relating to the field of materials technology. The preparation method of the PVDF membrane provided by this invention includes: coating a casting solution onto a substrate to form a liquid film; then, holding the carrier in an environment of 30-80°C for 1-5 seconds; subsequently, immersing the carrier in a coagulation bath to solidify it into a film; and finally, washing with water to obtain the PVDF membrane. This preparation method is environmentally friendly, simple, and low-cost. The obtained PVDF membrane has a dendritic-spherulitic alternating porous structure with controllable pore size, and can be used in biotechnology applications such as protein imprinting and sterilization filtration.

The application provides an anti-grass carp interleukin-21 monoclonal antibody and application, the monoclonal antibody is obtained by hybridoma cell strain with preservation number of CCTCC NO: C2022214, and the anti-grass carp interleukin-21 monoclonal antibody specifically recognizes grass carp interleukin-21 recombinant protein, and the anti-grass carp interleukin-21 monoclonal antibody can be applied in the preparation of reagent or kit for detecting grass carp interleukin-21 protein, and can also be applied in enzyme-linked immunosorbent assay, Western blotting, immunofluorescence assay, immunohistochemistry or flow cytometry detection;It can be applied to detect the production of interleukin-21 in grass carp tissue, and can also specifically detect the concentration of interleukin-21 in grass carp serum, provides parameter index for monitoring grass carp inflammation, can monitor the health status of grass carp in real time, thereby providing technical support for the sustainable development of grass carp breeding industry.

The application discloses a membrane strip incubation method for protein blotting and a detection method containing the same, and belongs to the technical field of immunoblotting. The membrane strip incubation method comprises the following steps: incubating a membrane strip by using a membrane strip incubation device, and placing the membrane strip into an incubation groove; adding buffer washing liquid and blocking liquid to the incubation groove, and blowing the liquid to flow to perform membrane blocking; adding a sample to be detected to the incubation groove, and blowing the liquid to flow to perform sample incubation; adding buffer washing liquid to the incubation groove, and blowing the liquid to flow to perform membrane washing; adding buffer washing liquid and enzyme combination to the incubation groove, and blowing the liquid to flow to perform enzyme combination; adding buffer washing liquid to the incubation groove, and blowing the liquid to flow to perform membrane washing; adding distilled water and color developing liquid to the incubation groove, shielding the membrane strip from light, and blowing the liquid to flow to perform light-proof color developing; adding distilled water to the incubation groove, and blowing the liquid to flow to perform color developing termination; and after each operation process is completed, liquid in the incubation groove is absorbed. The membrane strip incubation method ensures that the membrane strip is fully contacted with detection liquid and reacts.

A microfluidic Western blot method and system including a microfluidic western blot method for immunoassay of proteins, the method including introducing a sample including the proteins onto a chip; electrophoretically separating the proteins; binding the separated proteins to beads to form protein-attached beads, the beads being magnetic; flowing the protein-attached beads into a magnetic holding region; applying a magnetic field to the magnetic holding region to fix the protein-attached beads in place within the magnetic holding region; binding primary antibodies to target proteins on the protein-attached beads; binding secondary antibodies to the bound primary antibodies; and detecting the bound secondary antibodies.

This application provides an application of a reagent for quantitative detection of pyroglutamyl aminopeptidase in the diagnosis of rheumatoid arthritis (RA), where the reagent is a reagent for quantitative detection of pyroglutamyl aminopeptidase gene mRNA, a reagent for quantitative detection of pyroglutamyl aminopeptidase protein or a combination thereof. The quantitative detection is performed by enzyme-linked immunosorbent assay, Western blot, immunohistochemistry, a quantitative fluorescent polymerase chain reaction, mass spectrometry, or a combination thereof. A system for diagnosis of rheumatoid arthritis is also provided, including a sample collection module, a sample detection module and a data analysis module. The sample collection module is configured to collect a biological sample. The sample detection module is configured to quantitatively detect an expression level of pyroglutamyl aminopeptidase in vitro. The data analysis module is configured to compare a detection result with a diagnostic threshold, and generate a diagnosis report.

This application relates to the field of organic electrochemical transistors, particularly to their fabrication methods and applications. The organic electrochemical transistor includes a substrate layer, and a gate electrode, a source electrode, and a drain electrode disposed on the substrate layer. An organic semiconductor thin film is coated on the working area between the source and drain electrodes, and a biomodification material layer containing streptavidin is applied to the gate electrode. By setting the gate electrode, source electrode, and drain electrode on the substrate layer, and by applying a biomodification material layer containing streptavidin to the surface of the gate electrode, the streptavidin on the biomodification material layer can bind to biotin, thereby constructing an organic electrochemical transistor coated with an antigen-antibody trap. Changes in its relevant electrical parameters can be measured, and different concentrations of the target protein can be directly calculated. Therefore, compared to traditional protein blotting methods, this organic electrochemical transistor has the advantages of fast detection speed, high result sensitivity, fewer intermediate steps, and lower instrument costs.

The present application discloses the use of a compound targeting GPR183 in the preparation of a drug for treating acute myeloid leukemia. The compound is N-{6-[4-(2,4-dichlorobenzoyl)piperazin-1-yl]pyridin-3-yl}-N'-[4-(trifluoromethyl)phenyl]urea. The present application uses a computer virtual screening method to screen compounds effective against AML. Drug sensitivity experiments show that the compound has a killing effect on AML, and molecular docking analysis, RT-qPCR, and protein blotting prove that it can directly bind to GPR183, reduce the expression of GPR183 in AML cells, and inhibit AML cell proliferation. Therefore, the compound can be used as a compound targeting GPR183 in the preparation of a drug for treating acute myeloid leukemia.

The application provides a kit for identifying pig Y sperm and application thereof. Specifically, the kit takes pig membrane protein PPP4C or pig membrane protein CSNK2A2 as a molecular marker, tests protein expression amount by Western blotting or immunofluorescence, and identifies pig X and Y sperm according to the difference of the protein expression amount, and the specific identification standard is that the expression of pig membrane protein PPP4C or pig membrane protein CSNK2A2 in Y sperm is higher than that in X sperm. The application provides an effective scheme for realizing the identification of pig X and Y sperm by immunization, realizes the identification of pig X and Y sperm with low cost, high efficiency and simplicity, helps the sustainable development of animal husbandry, improves economic benefits, helps to alleviate the future food shortage problem, solves a series of animal welfare problems, and has important application value.

The present invention provides a pipetting mechanism in a protein blotting instrument, comprising a pipetting needle, a pipetting arm, and a cam, wherein the pipetting needle is fixed on the pipetting arm, the cam is in contact with the pipetting arm, a first rotating fulcrum is provided on the pipetting arm, the movement of the cam can drive the pipetting needle to rotate around the first rotating fulcrum, the pipetting needle can be used to absorb waste liquid in a reaction tank, and the pipetting mechanism also includes a supporting element, the supporting element is retractable or elastic, and the supporting element can allow the pipetting arm to always maintain contact with the cam. The structural design of the present invention allows the pipetting needle to remain stationary when in contact with the reaction tank, solving the problem of the pipetting needle hitting the needle and damaging the reaction tank in the prior art; the ordinary arc surface and the concentric arc surface on the cam have a smooth transition, so that there will be no excessive impact during the contact between the pipetting needle and the bottom of the reaction tank, protecting the reaction tank and ensuring the installation stability of the pipetting needle.

The invention provides a protein expression detection method based on an improved western blotting method, and the method comprises the following steps: carrying out pixel signal identification on a plurality of protein bands in a secondary antibody detection blotting membrane, and determining a detection limit value, background pixel intensity of a corresponding background signal in a set range of a target protein band, and a detection limit value of a target protein band; dividing the target pixel intensity by the background pixel intensity, generating a signal-to-noise ratio parameter of a target protein stripe, determining a pixel merging proportion of a pixel signal corresponding to the target protein stripe, identifying a pixel point location corresponding to the target protein stripe, performing pixel value merging on pixel points of the pixel point location corresponding to the target protein stripe according to the pixel merging proportion, and obtaining a pixel value merging result; and generating a western blot detection image of the target protein band, identifying lane size information of pixel points corresponding to the set protein band in the western blot detection image, and determining a protein expression detection result of the target protein band according to the linear relationship and the second lane size information. Therefore, the accuracy of protein quantitative detection is improved.

The utility model provides a western blot instrument feeding device. Relates to the technical field of protein analysis auxiliary tools. The western blot instrument feeding device comprises a portal frame, a rotating shaft is rotatably mounted at the top of the portal frame, a connecting disc is fixedly mounted at the top end of the rotating shaft, two supporting arms which are symmetrically arranged are fixedly mounted on the connecting disc, and first telescopic air cylinders are fixedly mounted at the tops of the two supporting arms; first n-shaped frames are fixedly installed on output shafts of the two first telescopic air cylinders correspondingly, second telescopic air cylinders are fixedly installed on the outer walls of the two sides of the two first n-shaped frames correspondingly, and output shafts of the four second telescopic air cylinders penetrate through the corresponding first n-shaped frames correspondingly and are in sliding connection with the corresponding first n-shaped frames correspondingly. According to the utility model, the time can be fully utilized, the storage bottle can be quickly replaced, and the analysis and detection efficiency is improved.