46 results about "Protein transfer" patented technology

Methods for transferring one or more proteins to a cell are disclosed. The protein or proteins to be transferred are in the form of a fusion protein, and contain at least one domain encoding for a protein or peptide having trans signaling and/or adhesion function. The fusion protein is transferred to a cell by binding to a lipidated protein, which has been incorporated into the cell membrane. In an additional aspect of the invention, methods of making fusion proteins having cis signaling capabilities, as well as the ability to bind with receptors on the cell's own surface, are provided. Fusion proteins incorporating GPI or a homing element, and a costimulator or inhibitor domain can also be directly transferred to the cell surface. Methods for using cells which have undergone protein transfer according to the present methods are also disclosed. This includes use in a cancer vaccine, use for treatment of cancer or autoimmune disease, and use in determining costimulator threshold levels.

The invention relates to the cell disruption of microbes and the preparation of the microbe proteins for mass spectrometric analysis. The cells of microbes from microcolonies are disrupted by physical or chemical means directly on the nutrient medium. The released proteins are then transferred to sample supports by direct contact with their contact surfaces; electrophoresis can be used for assistance. Once the the proteins are firmly adsorbed on the contact surfaces, they can be washed with water in order to remove substances which interfere with the ionization process. For analysis by matrix-assisted laser desorption (MALDI), the proteins are prepared on the contact surfaces of the sample supports with matrix substances to form MALDI samples; the sample supports are then introduced into a MALDI mass spectrometer for the acquisition of the mass spectra. The microbes are identified by similarity comparisons between the mass spectra of the microbe proteins and similarly obtained reference spectra.

The present invention relates to compositions having bioactive transfer proteins in stable carrier systems. Such systems are above all suitable for influencing/changing lipid cell structures by means of improved lipid exchange, while at the same time having great stability. In this connection, not only transfer proteins and lipid components, but also protective colloids are used, by means of which an impairment of the transfer effect of the proteins and a loss of stability can be avoided. The compositions are suitable for external topical use for cleansing, care, dermatological and/or therapeutic treatment of the skin, hair, or mucous membranes.

Disclosed herein are methods of protein quantification and normalization using haloalkylated tryptophan fluorescence. Complex protein samples, i.e., samples that each contain 1,000 or more distinct proteins, from diverse sources that do not have common protein profiles are treated with a halo-substituted organic compound (i.e. haloalkane) that reacts with tryptophan residues to form fluorescent products. Irradiation of the samples with ultraviolet light and the detection and quantification of the resultant fluorescent emissions from all proteins in each sample are then used to obtain comparative values for total protein content among the various samples. The values thus obtained are found to be valid indications of comparative total protein content, despite the fact that the tryptophan levels vary widely among the various proteins in any single sample and the samples, due to the diversity of their origins, tend to differ among themselves in the identities and relative amounts of the proteins that they contain. Protein samples are also normalized to correct for differences in sample dilution, sample loading, and protein transfer inconsistencies, by using stain-free detection of total protein in each of the samples, or detection of subsamples within each sample.

The invention discloses an electrochemical method for detecting the activity of protein acetyltransferase. P2 templated silver nanoparticles and cucurbit [8] urea are added into a P1 modified gold electrode and a detection system of a p300 protein, the activity of the p300 protein is detected by electrochemical measurement, and the protein transferase activity is obtained; the amino acid sequenceof P1 is 11-mercaptoundecanoic acid (MUA)-GGGFRGKGGKGLGKGGAKA; and the amino acid sequence of P2 is FGGGASLWWSEKL. p300 is used as a model, and the silver nanoparticles are synthesized by a signal template P2, and assembled with a sensing template P1 through a cavity, so that the sensitivity of detection is ensured, high design flexibility is achieved, and meanwhile, the selectivity of HATs is enhanced. The development of the simple and practical electrochemical method capable of detecting HATs activity is expected to help early diagnosis of major diseases.

The invention discloses a device for transferring and analyzing proteins in gel on line. The device comprises a three-electrode power supply, a capillary, a first buffering liquid tank, a second buffering liquid tank, a detector and a metal needle tube. During transferring, the metal needle tube and the zero electrode of the three-electrode power supply are connected, are sleeved on the sample introduction end of the capillary and are placed on the upper surface of the gel; the outlet end of the capillary and the positive electrode of the three-electrode power supply are placed in the first buffering liquid tank; and the negative electrode of the three-electrode power supply is connected to the gel. During analyzing, the zero electrode of the three-electrode power supply and the sample introduction end of the capillary are placed in the second buffering liquid tank; and the outlet end of the capillary and the positive electrode of the three-electrode power supply are still placed in the first buffering liquid tank. Compared with other protein transferring methods, the device can be used for transferring and detecting simply, fast, efficiently and one line, avoid the influence of dyeing on the transferring of a protein strip in SDS-PAGE, has important theory and practical values on gel protein transferring and has very wide practical application prospect.

Described herein is a device and method for translocating a protein through a nanopore and monitoring electronic changes caused by different amino acids in the protein. The device comprises a nanopore in a membrane, an amplifier for providing a voltage between the cis side and trans side of the membrane, and an NTP driven unfoldase which processed the protein to be translocated. The exemplified unfoldase is the ClpX unfoldase from E. coli.

The invention relates to a safety evaluation method of transferring Bt gene insect-resistant paddy for predator paederus fuscipes. The method comprises three parts which are three-level nutrition test, protein transfer rule and Tier-1 toxicity determination test, wherein each part evaluates the influence on the paederus fuscipes by the transferring Bt gene insect-resistant paddy through prey mediation, the approach and degree of the paederus fuscipes exposed to Bt protein, and the potential toxicity of the Bt protein expressed by the transferring Bt gene insect-resistant paddy to the paederus fuscipes, and finally ecological results to non-target organism by the transferring Bt gene insect-resistant paddy are comprehensively evaluated according to data on the three aspects. The evaluation method is systematic and scientific, and has important significance on perfecting the research on the influence of transferring Bt gene crop on non-target organisms and confirming the ecological safety of transferring Bt gene crop planting.

Kits, systems, and methods are provided for transferring biological macromolecules from an electrophoresis slab gel to a blotting membrane using conductive polymer electrodes.

The invention discloses a gradient gel and a preparation method thereof. The preparation method is characterized by comprising the following steps: respectively blending water, acrylamide separation gel stock solution, separation gel buffer solution, SDS (sodium dodecyl sulfate) and ammonium persulfate into two or more separation gels with concentration of 8-15% according to a certain proportion, quickly pouring the blended two or more separation gels in different concentrations into a glass plate crack in vertical slab electrophoresis from high to low concentration in order, adding water, adding spacer gel after the separation gel is solidified, and waiting the solidification of the spacer gel. According to the method, the electrophoresis speed is increased, the penetrability of protein molecules is enhanced, and the protein transfer rate and the protein transfer quality are improved.

In accordance with an embodiment of the present invention, there is provided a method of performing the western blot analytical technique, wherein the method comprises carrying out the following steps in the following order: a), pre-staining the proteins within a sample; b). separating the proteins using gel electrophoresis; c). analysing the separated proteins to determine the total protein load and/or at least one housekeeping protein load; d). transferring the proteins onto at least one membrane; e). probing the separated proteins to detect a target protein; and f). analysing the probed proteins to determine the target protein's molecular weight and/or load.

The invention relates to a preparation method of high-molecular-weight silver carp skin collagen peptide. The method includes the following steps of firstly, cleaning fresh silver carp skin to remove impurities, conducting degreasing through an alkali solution, removing impurity protein through an acid solution, and conducting rinsing to be nearly neutral; secondly, adding water to preprocessed silver carp skin, adjusting pH to be 3.5 to 4.5 and the temperature to be 40 DEG C to 45 DEG C, adding pepsase for an enzymatic hydrolysis reaction for the reaction time of 10 h to18 h, and keeping the pH and the temperature constant in the reaction process; thirdly, conducting ultra-low-temperature vacuum concentration on enzymatic hydrolysate of pepsase; fourthly, controlling the reaction conditions for a protein transfer reaction to obtain a large-molecule collagen peptide solution, and then conducting processing on the collagen peptide solution. By means of the method, protein synthesis is conducted on the basis of protein hydrolysis through the protein transfer reaction under the control conditions, large-molecule collagen peptide is obtained, molecular weight distribution is concentrated, raw materials and auxiliary materials are high in utilization rate, the process is simple, and processing cost is low.

The invention relates to a temperature controllable switching type nano-filtration membrane. The temperature controllable switching type nano-filtration membrane comprises a filtration membrane body and a carbon nanotube array which penetrates through the filtration membrane body, wherein the carbon nanotube array comprises a plurality of carbon nanotubes which are arrayed in parallel; the carbon nanotubes are grafted with temperature-sensitive macromolecular polymer material layers; the temperature-sensitive macromolecular polymer material layers are at a contracted or extending state along temperature changes, so that a pore diameter size of a channel of the temperature controllable switching type nano-filtration membrane is changed along the temperature changes; a preparation method comprises the following steps: taking the carbon nanotubes and poly(N-isopropyl acrylamide) as raw materials and preparing the nano-filtration membrane with temperature controllable ions and protein transferring by adopting a chemical vapor deposition method and a surface-initiated atom transfer radical polymerization method; the nano-filtration membrane has good hole permeability and sensitive environment response and can realize controllable transferring and filtering of inorganic ions and proteins under different temperature.

The invention discloses a high-efficiency protein transfer membrane buffer, which comprises the following components: the concentration of Tris is 10mM-100mM; the concentration of taurine is 10mM-200mM. Compared with the traditional membrane transfer buffer commonly used at present, the high-efficiency membrane transfer buffer of the present invention uses a taurine component with a stronger buffer capacity, which improves the buffer capacity of the buffer during electrophoresis, and is suitable for proteins with a wide range of molecular weights. Molecules have high membrane transfer efficiency, which can shorten the time for protein transfer to the membrane. At the same time, the heat generated by the buffer during electrophoresis is low, and there is no need to use ice packs to cool down, which simplifies the operation during membrane transfer. The concentration of methanol is lower, and a better transfer effect can be achieved at a concentration of 10%.

The invention is a high-throughput voltage screening crystallographic device and methodology that uses multiple micro wells and electric circuits capable of assaying different crystallization condition for the same or different proteins of interest at the same of different voltages under a humidity and temperature controlled environment. The protein is solubilized in a lipid matrix similar to the lipid composition of the protein in the native environment to ensure stability of the protein during crystallization. The invention provides a system and method where the protein is transferred to a lipid matrix that holds a resting membrane potential, which reduces the degree of conformational freedom of the protein. The invention overcomes the majority of the difficulties associated with vapor diffusion techniques and essentially reconstitutes the protein in its native lipid environment under “cuasi” physiological conditions.

A protein introduction method with which protein can be introduced into cells with excellent safety is provided. A target protein is supported on a carrier for protein introduction that is made from a clay mineral, and by adding this to cells it is possible to introduce the target protein into the cells. The clay mineral is preferably a layered clay mineral, and as the clay mineral it is possible to use montmorillonite, vermiculite, and illite, for example.

The invention discloses naphthyl urea-piperazine compounds and a preparation method and application thereof.The naphthyl urea-piperazine compounds can remarkably inhibit activation of cell signal transduction JAKs/STATs signals under the low dosage, and the experimental results of a cell proliferation experiment, an immunoblotting experiment, a cell cycle experiment, a protein transfer membrane transport experiment, a cell invasion and migration experiment and the like show that the naphthyl urea-piperazine compounds have the advantages that the naphthyl urea-piperazine compounds can be used for remarkably inhibiting activation of cell signal transduction JAKs/STATs signals; the compound can specifically inhibit JAK2 signal activation and expression of downstream STAT3, CyclinD1, CyclinB1, MMP9 and other target genes, induce blocking of a cell cycle G1 phase and cell apoptosis, can significantly inhibit proliferation of breast cancer, liver cancer, lung cancer, colon cancer, leukemia, lymphoma, multiple myeloma, retinoblastoma and other tumor cell strains, and can be used for preparing drugs for treating breast cancer, liver cancer, lung cancer, colon cancer, leukemia, lymphoma, multiple myeloma, retinoblastoma and the like. Therefore, the compound has a prospect of being developed into a targeted anti-cancer drug of a related target spot of a JAKs/STATs cell regulation signal transduction pathway.

The invention discloses a detection method of a collagen terminal peptide. The detection method comprises the following steps: (1) performing treatment and electrophoresis on a sample; (2) protein transfer printing; (3) closing and incubating: oscillating a transfer printing film obtained in step (2) into a closing solution; adding a collagen terminal peptide antibody to performing the primary antibody incubation for the transfer printing film, and then adding a secondary antibody to perform the incubation; and (4) performing the color developing reaction: adding the transfer printing film after the secondary antibody incubation of step (3) into a color developing solution to develop until a stripe appears, then adding distilled water to terminate the color developing reaction, and judgingthe type of the collagen terminal peptide in a to-be-detected sample according to the stripe. The invention provides the method which can effectively qualitatively and half-quantitatively detect thecontent of terminal peptide in exogenous collagen, and provides method evidence for the immunogenicity of an exogenous collagen-based biological product.