32 results about "Two-dimensional gel electrophoresis" patented technology

The present invention relates to protein markers for diagnosing stomach cancer and a diagnostic kit using the same, more precisely protein markers screened by two-dimensional gel electrophoresis and bioinformatics and a diagnostic kit using the same. The markers of the invention can be effectively used for diagnosing stomach cancer and evaluating the extent of progress of the cancer by confirming the expression levels of those marker proteins whose expressions differ in stomach cancer patients from in normal healthy people.

Disclosed are methods and apparatus for separation of biomolecules via two-dimensional gel electrophoresis, methods and apparatus for immunoblotting separated biomolecules, and methods for the use of biomolecules processed via the methods and apparatus of the present invention, including use in a clinical setting. The methods and apparatus for separation of biomolecules via two-dimensional gel comprises vertical agarose gel electrophoresis in the first dimension, and the electrophoresis of a novel non-denaturing 3-35% concave gradient polyacrylamide gel in the second dimension. This novel gel can be cast in a modified gel caster that can facilitate the pouring of multiple gels simultaneously. The methods and apparatus for immunblotting are useful with any type of immunoblotting, including Western blot, Northern blot, and Southern blot analyses. These methods and apparatus provide safe, efficient and cost-effective immunoblots, while facilitating the reduction of exposure to toxic or radioactive materials, as well as the disposal of those materials.

The invention provides a buildup method of a protein difference expression atlas when the committed differentiation of stem cells is induced by drugs, which detailedly establishes the protein difference expression atlas formed by that the stem cell is induced by the ICA and directionally differentiated to be a cardiac muscle cell; a model that the drug induces the stem cell to be directionally differentiated to be the cardiac muscle cell is established so as to prepare a two-dimensional electrophoresis protein sample; electrophoresis is gelled bidirectionally, images are collected and analyzed so as to confirm the difference protein. A comparison protein atlas is used for screening and identifying the difference expression protein when the ES cell is directionally differentiated to be the cardiac muscle cell by the inducing of the ICA; the difference expression protein is used as a drug effect target and applied to the preparation of a novel inducer which has high-efficiency and low-toxicity and is used for prompting the stem cell to be differentiated.

The invention relates to an extraction method of protein from animal tissues, in particular to an extraction method of total protein from a rumen epithelial tissue of a milk cow. The method comprises the steps of putting a rumen epithelial tissue sample of the milk cow in a precooled mortar, adding liquid nitrogen, grinding into powder, adding an acetone solution of precooled trichloroacetic acid, oscillating, eddying, uniformly mixing for overnight aging, centrifugally collecting sediment, freeze-drying the tissue sediment, adding lysate, incubating and eddying at intervals, and centrifugally collecting supernate, namely the total protein of the rumen epithelial tissue of the milk cow. For the total protein of the rumen epithelial tissue of the milk cow, extracted by the method, a total protein mixture can be separated by a two-dimensional gel electrophoresis technology according to an isoelectric point and molecular weight of the protein, the relative abundance of the protein is acquired, the isoelectric point, the molecular weight and a relative transcript level of each protein can be clearly and visually shown in a gel atlas, and the method has the advantages of more obvious integrity and visualization in comparison with the prior arts such as a high efficiency liquid chromatography separation method, an enzyme-linked immunosorbent assay and immunoblotting.

The invention discloses a preparation method of a micro-fluidic chip interface for two-dimensional protein electrophoretic separation, and relates to a micro-fluidic chip. The method comprises the following steps of: preparing polyacrylamide gel in a vertical channel of the micro-fluidic chip and filling a buffering aqueous solution; introducing an organic phase through a horizontal channel of the micro-fluidic chip and filling the horizontal channel; introducing an organic phase into which tetraisopropyl titanate is dissolved and filling the horizontal channel; draining an organic solution of tetraisopropyl titanate out of the horizontal channel, and cleaning the horizontal channel; adding an isoelectric focusing buffer solution into which proteins and amphoteric electrolytes are dissolved into the horizontal channel, applying electric fields to the two ends of the horizontal channel for performing isoelectric focusing, and realizing first-dimension separation of proteins in the horizontal channel according to respective different isoelectric points; and after finishing isoelectric focusing, applying electric fields to the two ends of the horizontal channel for performing second-dimension gel electrophoresis, making micropores on a TiO2 film pass through an electric field, and making the proteins enter a second-dimension channel by passing through the micropores on the TiO2 film under the action of the electric field for performing two-dimension separation.

The invention discloses an identification method of intracellular and extracellular proteomes of penicillium during fermentation. The identification method comprises the following steps of: (1) extracting intracytoplasmic proteins; (2) extracting extracytoplasmic proteins; (3) carrying out two-dimensional gel electrophoresis; (4) analyzing a two-dimensional gel image; and (5) identifying the proteins by using MALDI (Matrix-Assisted Laser Desorption Ionization)-TOF (Time-Of-Flight)-MS (Mass Spectrometry). The protein level closely related to penicillin synthesis in a penicillin fermentation process, and a relation between the protein level and the penicillin production level can be found quickly and accurately in high flux by using the identification method, differentially expressed proteins are found out and provide a target spot for further modifying a penicillium metabolic path, thus a certain theory is provided for obtaining high production strains and improving the yield of penicillin, and a new method and a concept are provided for modifying other industrial antibiotic strains.

The invention relates to an application of human derived HSF2 as the specific diagnosis molecular marker of ulcerative colitis, and belongs to the technical field of biomedicine. The technical scheme comprises three parts: (1) detecting differential proteins in serums of UC patients and healthy volunteers through a proteomics two-dimensional gel electrophoresis technology and a mass spectrum technology; (2) detecting the HSF2 expression level in colonic mucosa of UC patients in different degrees through an immunohistochemical method; (3) detecting the HSF2 expression level in colonic mucosa of UC patients in different degrees through a real-time quantitative PCR technology and a western blotting technology. 40 UC patient serums and 40 healthy volunteer serums have been compared by the method mentioned above, and the results show that the expression of HSF2 in the UC patient serums prominently increases. Moreover, an immunohistochemical method, a real-time quantitative PCR technology and a western blotting technology are adopted to detect the HSF2 expression in the colonic mucosa of UC patients, and the results show that the HSF2 expression increases as the UC disease goes worse. Thus, the HSF2 can be used as a molecular marker to specifically diagnosing ulcerative colitis.

The invention discloses a protein complex or compound three-dimensional gel electrophoresis separating method. The method comprises the following steps: 1) separating the protein complex by non-denatured polyacrylamide gel electrophoresis method and then dyeing the protein complex; 2) chopping the gel strips containing the target protein complex or compound obtained in step 1), and decoloring and dehydrating; and 3) soaking the gel strips processed in step 2) by protein denaturation solution, adequately pulverizing and directly performing separation by two-dimensional gel electrophoresis method. The method in the invention dispenses with protein electroelution and protein deposition operating procedures in traditional protein complex or compound three-dimensional gel electrophoresis method, thus avoiding protein loss and protein in vitro chemical decoration; and the method in the invention is simple to operate and needs no additional equipment. Proved by experiments, the method in the invention achieves good protein complex or compound electrophoresis separating effect.

The invention relates to a method for utilizing a proteomic technology to screen a drug action target position of cyclosporine A in an immunized T cell. According to the method for screening the drug action target position of cyclosporine A in the immunized T cell, the proteomic technology is used for screening the drug action target position of cyclosporine A in the immunized T cell and the method comprises following steps: step 1) establishing sample groups of the action of cyclosporine A immunizing the T cell; step 2) preparing two-dimensional electrophoresis protein samples of the immunized T cell; step 3) carrying out two-dimensional gel electrophoresis; step 4) collecting images and analyzing to obtain the result. The method for screening the drug action target position of cyclosporine A in the immunized T cell, disclosed by the invention, is clear and definite in operation through, and true and reliable in result, overcomes the defects that with adoption of the conventional screening method, the screening result is high in error and low in accuracy as the analysis data are huge and complicated, and provides the detection result with pharmacologic significance and guiding function for clinical medication.

The invention provides a biomarker method for identifying the existence form of human pituitary tissue lactation hormone protein. The method comprises the following steps of collecting pituitary adenoma and normal pituitary tissue samples, respectively extracting the tissue proteins, scanning a visual PVDF membrane and bidirectional gel into a digital image through bidirectional gel electrophoresis and western blot silver staining, digesting and purifying the corresponding two-way gel protein spots, and identifying the difference of the existing form proportions of different pituitary tissue lactation hormone proteins of different pituitary tumor sub-types through mass spectrum identification and bioinformatics analysis. According to the present invention, the existence forms of the human pituitary tissue lactation hormone protein and the differential expression profiles of the prolactin proteins in different pituitary tumors are identified, so that the development of drugs for blocking prolactin signal channels for clinical treatment is facilitated.

The invention relates to a method for utilizing a proteomic technology to screen a drug action target position of mizoribine in an immunized T cell. According to the method for screening the drug action target position of mizoribine in the immunized T cell, the proteomic technology is used for screening the drug action target position of mizoribine in the immunized T cell and the method comprises the following steps: step 1) establishing sample groups of the action of mizoribine immunizing the T cell; step 2) preparing two-dimensional electrophoresis protein samples of the immunized T cell; step 3) carrying out two-dimensional gel electrophoresis; step 4) collecting images and analyzing to obtain the result. The method for screening the drug action target position of mizoribine in the immunized T cell, disclosed by the invention, is clear and definite in operation through, and true and reliable in result, overcomes the defects that with adoption of the conventional screening method, the screening result is high in error and low in accuracy as the analysis data are huge and complicated, and provides the detection result with pharmacologic significance and guiding function for clinical medication.

The invention belongs to biological gene engineering field, especially relates to an Aegilops tauschii high molecular weight glutelin 1Dy12.1#+[t] gene and its uses in biological improvement of wheat quality based on the gene. Glutelin gene is mainly separated using library establishment method and PCR method. In the invention, a new high molecular weight glutelin HMW-GS12.1#+[t] and its gene are separated from Aegilops tauschii using allele specific PCR, single-direction or two dimensional gel electrophoresis and capillary electrophoresis. The gene sequence has high homologue with quality gene 1Dy10, can work in wheat quality improvement.