53 results about "Gel electrophoresis of proteins" patented technology

The present invention relates to a lab-on-chip substrate, comprising a resin having a silicon content of 10% or less by weight as its base material and a hydrophilic polymer covalently bound onto the surface thereof by high-energy ray irradiation, and in particular, to a protein-processing chip. The present invention provides a lab-on-chip substrate resistant to washing and usable for an extended period of time without adsorption of proteins on the base material surface, i.e., a protein electrophoretic polymeric chip having a microchannel allowing high-accuracy analysis of trace amounts of proteins because of reduction in the amount of detection noise.

The invention discloses a method for gel electrophoresis separation of serum, wherein sectionalized polyacrylamide gels with three different concentrations are used as electrophoresis carrier for forming temperature gradient in the electrophoresis tank, so as to achieve synchronous electrophoretic separation for serum lipoprotein and its subcomponents, wherein the concentrations of the polyacrylamide gel being separation gel 1 3.0%, separation gel 2 3.6%, separation gel 3 7.0%.

Disclosed is a leak-proof gel casting device for a vertical protein electrophoresis system which includes a gel caster which has a plurality of compartments and a sealing pad which is inserted to the compartments of the gel cater, and is formed to wrap around both sides and an edge of a lower side of a plate set. The present invention with a simple structure is capable of being manufactured at a low cost, does not require frequent replacement because of its long-term durability, and is highly effective in preventing the leakage of the gel placed in the gap between the plates.

The invention provides a preparation method of a water-soluble human collagen VI polypeptide, which comprises the following steps: linking coding genes of the human collagen VI polypeptide into a cloning vector, transforming Escherichia coli, constructing engineering bacteria, inducing the engineering bacteria to express the human collagen VI polypeptide, and respectively establishing an engineering bacteria fermentation process and a target protein purification process. The protein electrophoretic analysis and activity verification indicate that the molecular weight of the human collagen VI polypeptide is 46kDa, and the human collagen VI polypeptide is soluble in supernatant, has favorable water solubility and exists in the form of a monomer or polymer. The human collagen VI polypeptide in vitro can protect cells from ultraviolet injury, and has the functions of diminishing inflammation and moistening. The invention realizes a large-scale gene engineering preparation method of a water-soluble human collagen VI polypeptide.

A prefabricated colloid used for electrophoretic analysis of protein contains polyacrylamide gel as main component, storage liquid (pH 6.5), neutral Bis-Tris buffering system (pH 6.5), ammonium persulfate, neutral MOPS-SDS electrophorectic buffering liquid and EDTA. Its advantages are high stability and easy storage and application.

The invention discloses a method utilizing internal peptide inverse splicing to prepare protein molecular weight standard, which includes: selecting a section of known protein X with a certain molecular weight, selecting two internal peptides A and B not compatible to each other, and then respectively decompounding internal peptides A and B into amino terminal (N terminal) and carboxyl terminal (C terminal), namely An, Ac, Bn, Bc; and then using them to consist five recombinant proteins, namely an initial point protein (X-An), an intermediate section 1 protein(Ac-X-Bn), an intermediate section 2 protein(Bc-X-An), an end point 1 protein(Ac-X), and an end point 2 protein(Bc-X); obtaining protein molecular weight standard with multiple molecular weight distribution by mutually combining, connecting and overlapping the five proteins.

The invention relates to a qualitative detection method capable of distinguishing cow milk doped in human milk. The qualitative detection method is a protein gel electrophoresis method. The electrophoresis method comprises the following steps of: putting a prepared gel plate into an electrophoresis slot, adding an electrode buffer solution, performing pre-electrophoresis under 30 to 120 volts for 15 to 60 minutes, injecting the treated test sample solution into a gel sample application hole, performing electrophoresis under 50 to 200 volts until a bromophenol blue indicator is electrophoresized to the bottom of the gel, and finally taking out the gel and fixing in a fixing solution, dyeing in a dyeing solution and decoloring in a decoloring solution so as to obtain a protein electrophoresis pattern; and comparing the sample electrophoresis pattern with a characteristic protein band in the electrophoresis pattern of pure human milk and pure cow milk prepared under the same condition so as to determine whether liquid cow milk or milk powder reconstituted milk is doped into the sample milk. In the method, only one gel electrophoresis apparatus and a conventional chemical reagent are needed, the operation is simple and the result is reliable; and the cow milk or a product of the cow milk doped into the human milk can be detected.

The invention relates to application of lactobacillus plantarum subspecies and lactobacillus casei 1 in horseradish tree leaf fermentation, and belongs to the technical field of dairy processing. Seven strains of lactic acid bacteria are domesticated through the gradient culture method of gradually increasing the proportion of horseradish tree liquid in skim milk so that the adaptability of the lactic acid bacteria can be enhanced and the growth situations of domesticated leavening agents can be studied. The free amino acid content of the horseradish tree liquid fermented through the seven strains of lactic acid bacteria is studied, four strains good in degradation are screened out, changes of protein electrophoretic bands and peptide substances of the strains are analyzed through the SDS-PAGE technology and the HPLC technology, and the degradation effect of the leavening agents on horseradish tree proteins is studied. Finally, the lactobacillus plantarum subspecies and the lactobacillus casei 1 which are screened out serve as the leavening agents to be applied to the horseradish tree leaf fermentation. The application has the advantages that the horseradish tree leaf proteins can be well degraded, and the protein utilization rate is remarkably increased; horseradish tree leaves are weakened in bitter and spicy taste and improved in taste after being fermented; the horseradish tree industrial development can be easily promoted.

The invention discloses a method for separating antibacterial proteins from bacillus pumilus E14. The method comprises the following steps: preparing a zymophyte solution through fermentation production, and centrifuging to prepare supernatant; precipitating by using ammonium sulfate, resuspending the obtained precipitate by using a buffer solution to obtain resuspension; separating through an ion exchange method, collecting an elution peak, concentrating, dialyzing, and obtaining an active ingredient separation peak; and measuring the molecular weight of the antibacterial proteins separated from the bacillus pumilus E14 to be between 97.2 and 116.0kD through antibacterial activity detection and protein electrophoresis detection. The method is simple and reasonable in design and convenient to operate, and proteins with an effect of resisting Vibrio harveyi ((i) Vibrio( / i) (i) harveyi( / i) can be separated according to the method.

The invention relates to a Chinese date honey characteristic protein and isolation and purification thereof; firstly, eight single-flower honey is subjected to SDS-PAGE, a result finds out that the Chinese date honey has one protein spectral band at about 17 ku while protein electrophoresis spectrograms of other honeys do not contain the protein spectral band at about 17 ku; in addition, the molecular weight of the protein has quite large difference from the molecular weights of other protein spectral bands, so as to facilitate separation. Therefore, the protein can be used as the characteristic protein of the Chinese date honey different from other honeys. For making the molecular weight and structure information of the protein clear, an LC-Chip / ESI-QTOF-MS method is used for identification. The operation is simple, the separation effect is good, the feasibility is high, and the new method is provided for true and false identification of the Chinese date honey.

The invention relates to a spirulina development and application technology and aims to provide a method for distinguishing the degrees of production traits of a spirulina strain. The method provided by the invention comprises the following steps of: extracting water-soluble protein of spirulina cells with an tris-HCl extracting solution, separating a protein sample by carrying out sodium dodecyl sulphate-polyacrylamide gel electrophoresis, then detecting the light intensity value of a protein band at a 102kD site of a protein electrophoretogram, and constructing a hierarchical diagram among algae plants according to the light intensity value; if the light intensity value of the protein band of a candidate strain at the 102kD site is more than 140 and the candidate strain and a known strain with good production traits get together, indicating that the candidate strain has good production traits and applicable to large-scale cultivation production; and if the light intensity value of the protein band of the candidate strain at the 102kD site is less than 40 and the candidate strain and a known strain with poor production traits get together, indicating that the candidate strain has bad production traits and is not applicable to large-scale cultivation production. The method provided by the invention is simple, efficient, reliable and low in cost, and no complex test and analysis such as nucleic acid sequencing and bioinformatics comparison is carried out, so that the method provided by the invention is applicable to large-scale high throughput screening.

This invention provides a reagent for protein electrophoresis, an electrophoresis gel or buffer composition containing the reagent, and a protein separation method and an electrophoresis kit using the reagent and the composition.

The invention relates to the technique of development and application of spirulina and aims at providing a method for detecting production trait goodness and badness of spirulina strains. According to the method, a Tris-HC1 extracting solution is utilized for extraction so as to obtain water-soluble protein of spirulina cells; protein samples are separated by using sodium dodecyl sulfate-polyacrylamide gel electrophoresis, subsequently the optical density value of a protein zone at 102kD of a protein electrophoresis diagram is detected, and a dendrogram of the spirulina plants is established according to the value; if the optical density value of a candidate strain at the protein zone at the 102kD is larger than 140 and the candidate strain is gathered together with a known stain with good production nature, the stain is good in production nature and is applicable to large-scale cultivation and production; and if the optical intensity value of the protein zone at 102kD is less than 40 and the candidate strain is gathered together with known strains with poor production nature, the stain is poor in production nature and is not applicable to large-scale cultivation and production. The method is simple, efficient, reliable and low in cost, and complex tests and analysis on nucleic acid sequencing and bioinformatics comparison do not need to be carried out, therefore, the method is applicable to large-scale and high flux separation.

The invention discloses a protein isoelectric focusing electrophoresis method and glue preparation device, and belongs to the technical field of bioseparation detection. The method comprises the stepsof glue preparation, glue filling, pre-electrophoresis, electrophoresis, gel fixing, decoloration, dyeing and the like. According to the method, the feeding mode of a sample and voltage and electrophoresis time of pre-electrophoresis and electrophoresis are changed, so that obtained protein electrophoresis bands are clear, good in separation degree and do not diffuse or trail, the problem that inthe prior art, the electrophoresis time is long, and protein diffuses and trails is effectively solved, and the glue preparation success rate is greatly increased. The glue preparation device comprises a glue preparation support, a fixed frame, glue preparation glass, a gasket, an anti-leakage pad and other components. By improving the structure of the glue preparation device, the consumption ofa reagent is reduced, glue filling is fast, no bubble is generated, glue gathering is even, the glue preparation success rate is greatly increased, and meanwhile the inspection cost is reduced.

The invention belongs to a wheat radiation breeding method, in particular to a 60Co-Gamma radiation method for breeding a new wheat high-molecular-weight glutenin subunit line. Reports on the 60Co-Gamma ray breeding of a new wheat high-molecular-weight glutenin subunit mutant line in the prior art are not seen. The method includes the following steps: wheat seeds are irradiated by 60Co-Gamma rays for ten minutes, and the dosage rate is 200Gy; a first mutagenized generation is produced; seeds of a second mutagenized generation are produced; individuals of the M2 generation are grown and harvested; the wheat glutenin of the M2 generation is extracted by the half-seed method, and protein electrophoresis is carried out; half seeds are grown, and a third mutagenized generation is harvested; the wheat glutenin of the M3 generation is extracted; M3 protein electrophoresis is carried out; and the new mutant line with different number of high-molecular-weight glutenin subunit bands or high-molecular-weight glutenin subunit bands at different positions is determined by comparing with the parents. The method has the advantages that: the method breeds the new wheat high-molecular-weight glutenin subunit mutant line by combining 60Co-Gamma ray radiation with the electrophoresis and combining the field planting with indoor electrophoresis screening; and the result is accurate, scientific and reliable.

The invention discloses a method for acquiring a two-dimensional electrophoretogram of nectar protein of liriodendron tulipifera by the aid of a two-dimensional electrophoresis system. After a nectar sample is filtered by an ultramicro filter membrane, the nectar protein is concentrated and purified by a protein ultrafiltration and concentration purification column, the concentration of the purified and concentrated nectar protein is detected with a Bradford method, the molecular weight distribution range of the nectar protein of the liriodendron tulipifera is pre-detected through one-dimensional lauryl sodium sulfate polyacrylamide gel electrophoresis, finally, proper standard protein and a gel strip with a proper length and pH range are selected according to a one-dimensional electrophoresis result, the nectar protein of the liriodendron tulipifera is separated with the two-dimensional electrophoresis system, and the two-dimensional electrophoretogram is acquired. The method has the characteristics that the operation is simple, the protein extraction rate is high and few interfering substances exist and is suitable for special materials such as liquid samples, the high-quality two-dimensional gel electrophoretogram with high resolution ratio, clear protein points, larger numbers, uniform distribution and clear background can be obtained, and the experiment repeatability and the stability are good.

The invention provides a protein electrophoresis method and relates to the field of protein analysis. The invention particularly relates to preparation of single-layer polyacrylamide gel and a protein electrophoresis analytic method therewith. The preparation of the single-layer polyacrylamide gel is simple in processes and is low in time requirement. The method is excellent in a separation effect on proteins being 4-100 kDa in molecular weight.

The invention discloses an electrophoresis apparatus with a concentration function and relates to the technical field of electrophoresis apparatuses. The electrophoresis apparatus comprises an electrophoresis apparatus shell, a central electrode, a peripheral electrode, a medium layer and an electrophoresis power supply, wherein the central electrode is mounted at the central position in the electrophoresis apparatus shell; the peripheral electrode is mounted in the electrophoresis apparatus shell and surrounds the central electrode; the medium layer is arranged between the central electrode and the peripheral electrode and can realize protein electrophoresis; the electrophoresis power supply is connected with the central electrode and the peripheral electrode respectively and is used for forming an electric field capable of separating proteins and concentrating separated components after the central electrode and the peripheral electrode are electrified by the electrophoresis power supply. According to the electrophoresis apparatus with the concentration function, a cylindrical electrode and an annular electrode are used and the electric field capable of separating the proteins and concentrating the separated components is formed, so that the separated proteins have the concentration function and user experience is improved.

A gel electrophoresis apparatus for single cell gel electrophoresis, including a chamber for receiving a gel electrophoresis buffer, a functional cover for closing the chamber, at least one pair of electrodes for generating a homogeneous electric field in the chamber and at least one retaining element for receiving and positioning at least one support plate. The at least one retaining element positions the at least one support plate in the homogeneous electric field generated by the at least one pair of electrodes.

The invention discloses a method for detecting burkholderia cepacia flora on fruits or vegetables and identifying seeds. The method comprises the following steps: separating flora on sick-healthy borderlines, extracting DNA, utilizing specific primers to perform PCR amplification, separating amplification products through gel electrophoresis, use an indicator to develop color and identifying whether a strain belongs to burkholderia cepacia flora according to band situations; performing enzyme digestion on PCR products of each strain, obtaining a restriction enzyme digestion map and preliminarily determining seed of each strain; utilizing the specific primers of corresponding seeds to perform PCR amplification and judging the seeds of the strains after gel electrophoresis separation according to the band situations; and sequencing last PCR products of the strain if gel shows no band, obtaining a recA sequence, constructing a molecular phylogenetic tree and determining the seed of the strain. The method can accurately detect the existence of Bcc bacteria on fruits and vegetable with pathological changes and confirm every seed in Bcc flora in order to take effective measures to dangerous bacteria.

The invention discloses a reagent for coloration of protein in gel electrophoresis as well as a using method and application. The reagent comprises a first component and a second component, wherein the first component is the solution formed by dissolving the compound with isothiocyanic acid groups into a solvent for protecting the isothiocyanic acid groups; the second component is the alkaline loading buffer which does not contain amino groups; the using method comprises the steps as follows: mixing the first component and the second component to form treating fluid and mixing the treating fluid with a to-be-tested protein sample, or mixing the second component with the to-be-tested protein sample and then adding the first component, and heating to form the chromoprotein sample solution. According to the invention, the reagent can be used in gel electrophoresis, and has the advantages of simplicity and easiness in use, reliability in effect, low usage amount, easiness in production and processing, wide application range, no pollution, time conservation and labor conversation, capabilities of performing real-time observation and being used for pre-colorating of the protein in gel electrophoresis, and the like, and a protein electrophoresis pattern has no changes which can be observed.

The invention provides a method for improving experiment accuracy by an isoelectric focusing method of two-dimensional protein electrophoresis, in particular a method for calculating electric energy needed by salt and protein focusing by measuring electric conductivity of a protein object to be measured. The method is characterized in that: electric energy needed by the protein focusing and salt mobility is calculated through a group of formulas in environments of different electric conductivities of the salt, the protein weight, the pH gradient gel length and the range of the pH value respectively, proper electric energy of the experiment of the individual protein object to be measured can be provided in the isoelectric focusing method so as to present an optimal protein focusing result,and the influence of colloidally protein focusing due to insufficient or over-high electric energy is avoided.

The invention discloses a method for preparing a capillary inner wall coating, and aims to provide the method for preparing the capillary inner wall coating which has the characteristics of quick, simple and convenient operation, low material cost and stable product performance. The method comprises the following steps of: rinsing a quartz capillary with the solution of citric acid; dissolving polyethylene glycol 2000 into the solution of the citric acid which has the pH of 2.2 and the concentration of 50mM and serves as a medium to prepare 0.8 to 10 percent coating buffer solution; dissolving a protein to be detected into the coating buffer solution; and performing electrophoretic separation on the protein, simultaneously finishing preparing the capillary inner wall coating. The method is suitable for the technical field of protein electrophoretic separation analysis.

The invention provides a single-cell protein quantitative analysis method based on an electrophoresis technology, which comprises the following steps: 1) carrying out physical separation through a micropore array chip consisting of single-cell separation chips to obtain single cells, and carrying out in-situ lysis on the single cells to carry out protein electrophoretic separation; 2) transferring a protein strip obtained by electrophoretic separation to an inverse opal structure photonic crystal film; 3) incubating an SERS nano-label developer and the inverse opal structure photonic crystal transfer printing film, and developing protein on a strip through an SERS imaging technology; and 4) performing Raman spectrum analysis, and calculating the concentration of each protein according to the Raman characteristic peak intensity of the nano-label. The method has the beneficial effects that a novel single-cell protein quantitative analysis method is constructed by combining a protein electrophoresis technology, SERS (Surface Enhanced Raman Scattering) and an ordered structure photonic crystal film, and high-sensitivity and wide-linearity quantitative detection requirements can be met at the same time.

The invention provides an antimicrobial experimenting method capable of determining an antimicrobial protein fragment molecular weight, and relates to the technical field of microorganism. After a reagent, separation gel and spacer gel are prepared, protein for antimicrobial experimenting is subjected to protein electrophoresis, an obtained protein electrophoresis film is subjected to decolorization and dyeing, and after the size of a protein molecular weight is observed through imaging, the protein electrophoresis film is used for the antimicrobial experiment. After the protein electrophoresis film is placed in a PBS buffer solution and balanced for 10 minutes, the protein electrophoresis film is placed in an agar culture dish. A bacterium solution in a logarithmic phase is taken out and added into a nutrient agar culture medium at a temperature of 50 DEG C, and the bacterium solution with a volume concentration of one percent is obtained. The bacterium solution is taken out and poured into the culture dish containing the decolorized protein electrophoresis film, the growth situation of thallus in the culture medium is observed, and the size of protein fragment molecular weight which has the good antimicrobial effect is determined. The antimicrobial experimenting method used for determining the size of the antimicrobial protein fragment molecular weight with the antimicrobial effect has the advantages of being easy to operate, accurate, rapid and cheap. The antimicrobial experimenting method has a wide application prospect.

The invention belongs to the technical field of virus antibody preparation, and particularly relates to a preparation method of a rice dwarf virus antibody. The invention provides the preparation method of the rice dwarf virus antibody, which solves the problem that macromolecular target proteins are generally not easy to induce. The preparation method comprises the following steps: (1) inoculating a positive strain in two tubes of LB fluid mediums containing penbritin and chloramphenicol according to a ratio of 1 to 100, shaking for 3 to 4 h, and performing induced culture for 4 to 5 h, wherein IPTG is not added in another test tube for contrast; taking bacterial fluid, collecting thalli, and putting into Coomassie brilliant blue staining fluid after the thalli are subjected to gel electrophoresis separation; and (2) inducing 100 ml of the bacterial fluid, collecting thalli, performing gel electrophoresis on target proteins, performing KCL pre-cooling on gel subjected to electrophoresis to obtain a milk white protein band, cutting and recycling the target protein band with a blade, adding the freund's adjuvant, grinding to obtain emulsified proteins, and storing the emulsified proteins for later use.

The invention belongs to the field of bioengineering, and particularly relates to a method for determining the minimum entrapment binding ratio in a polypeptide and siRNA co-assembly. The invention discloses a method for determining the minimum entrapment binding ratio in a polypeptide and siRNA co-assembly. The method comprises the following steps: S1, preparing 2% agarose gel; s2, preparing the CPP-siRNA nano particles; s3, target polypeptide is added, an electrophoresis experiment is carried out, and a gel imager is used for imaging; and observing a map in an imaging result by naked eyes, selecting a group with no tailing of a protein electrophoresis track in the image, and taking the minimum binding ratio in the group without tailing as the minimum entrapment binding ratio. The invention discovers a method for determining the minimum entrapment binding ratio of the positively charged nanoparticles formed by self-assembly of the polypeptide and the siRNA, discovers several polypeptide sequences suitable for the method, and fills the blank in the prior art. The method is simple to operate, and the judgment method is very visual and easy to operate.