67 results about "GTP-sepharose" patented technology

The invention discloses a preparation method of magnetic sepharose gel microspheres. The preparation method comprises the following steps: (1) preparing ferrous powder with scattered active agent: adding the ferrous powder into surface active agent water solution, heating and mechanically stirring; washing and drying the obtained product to obtain the ferrous powder with scattered active agent; and (2) preparing the magnetic sepharose gel microspheres: dissolving an emulsifying agent span80 into liquid paraffin for preheating; taking sepharose gel and the ferrous powder with scattered active agent, adding secondary distilled water to prepare sepharose solution, conducting ultrasonic scattering and heating and dissolving; adding the dissolved sepharose solution into span80-containing liquid paraffin solution; after the reaction is finished, cooling a sample and conducting slow and mechanical stirring; separating the sample, and washing to obtain the magnetic sepharose gel microspheres. By adopting a reversed phase suspension embedding method, the prepared magnetic sepharose gel microspheres are strong in magnetic responsibility, and have a good effect on the separation and purification of protein.

The invention discloses a method for detecting the cytotoxicity of a to-be-tested medicine to a target cell and a cell chip specially used by the method. The cell chip provided by the invention comprises a supporting medium and a sepharose gel film attached to the upper part of the supporting medium, wherein a plurality of microtraps are distributed in the sepharose gel film, and the size of each microtrap is matched with that of the individual target cell; and the individual target cell is attached to the inside of each microtrap. The experiment proves that the cell chip provided by the invention can detect the cytotoxicity of the to-be-tested medicine to the cell and/or detect the gene damage of the to-be-tested medicine to the cell and/or detect the influences of the to-be-tested medicine to the expression of interest protein in the cell and/or screen high-content drugs having effects for the cells. The cell chip provided by the invention has important application values.

The invention relates to a DEAE dextran-modified agarose gel-based chromatography medium and a preparation method and application thereof. The method includes the steps that first, the molecular weight of DEAE Dextran for grafting is selected; next, the concentration of a DEAE Dextran aqueous solution is controlled; then, the time of diffusion before DEAE Dextran coupling is determined; finally, the reaction time of coupled DEAE Dextran and the concentration and volume of sodium hydroxide are determined. The DEAE dextran-modified agarose gel-based chromatography medium has a strong adsorption property for protein under different modification densities, shows a high adsorption capacity and a high adsorption rate and improves separation efficiency. The medium is convenient to clean and degerm, easy to regenerate and good in biocompatibility. The preparation method is simple and harmfulless and has broad application prospects in efficient and rapid separation and purification of protein.

The invention relates to the field of gene engineering, and discloses a construction and an expression for two engineering bacteria of two human glucagon-like peptide-1 analogues, and a preparation method for the objective peptides. The objective peptides are formed by inserting objective sequence after double digestions, adding His labels at the front ends of fusion proteins, obtaining objective peptides by cutting the fusion proteins with acid via gaining the fusion proteins, and after an isoelectric precipitation, obtaining the objective peptides again through an affinity chromatography of a Ni agarose gel after an isoelectric precipitation. Freeze-dried objective peptides are dissolved in a PBS buffer solution, oral glucose tolerance test and determination of serum insulin concentration are carried out on normal ICR mice. The two analogues are modified peptides of human glucagon-like peptide-1. Experiments demonstrate that both the two peptides have effects for reducing blood glucose and can be used for treating type 2 diabetes mellitus. Furthermore, the invention also provides the preparation method for the peptides.

The invention relates to a bilateral molecular marker identification technology of a melon wilt resistant variety. By the utilization of polymerase chain reaction and bilateral molecular markers ML430 and MR296 which are closely linked to a melon wilt resistance gene, specific DNA fragments are amplified. The size differences of amplified DNA fragments between a resistant variety and a susceptible variety can be displayed on different locations of a sepharose gel by electrophoretic technique. And the sizes and locations are compared with that of DNA fragments amplified from a standard resistant variety and a standard susceptible variety. Two varieties which have the same size or lie in the same location belong to the variety of the same type. When banding patterns of the two molecular markers both are resistance type, the variety is the resistant variety. The method provided by the invention has advantages of convenient sampling, precise identification, and no influence by plant growth and development and weather conditions. The technology provided by the invention is mainly used for the screening and identification of wilt resistance variety resources, and can also be used for molecular marker assisted breeding of melon.

The invention discloses a multi-purpose separation medium and a preparation method thereof, wherein, tetrazole is the functional group of the medium. More particularly, a separation medium with novel structure is prepared by linking the tetrazole on the surface of silica gel, dextran and agarose gel separation group. The invention has dual purposes of being applied to ion-exchange chromatography and metal chelate chromatography. When the medium is applied to the ion-exchange separation of proteins, the separation of proteins is highly selective, easy to regenerate and quick to separate and both the quality and recovery rate of the activity of the proteins are high. When the medium is applied to the metal chelate chromatography of proteins, the separation of proteins is highly selective, small in metal ion loss and quick to separate and both the quality and recovery rate of activity of the proteins are high. The invention can be applied to the quick separation and purification of genetic engineering products and plasma proteins.

The invention discloses a preparation method of an ulva fasciata delile sulfated polysaccharide with anti-tumor activity. The preparation method comprises the following steps of crushing and ultrasonically extracting ulva fasciata delile by water to prepare a crude polysaccharide through alcohol precipitation; carrying out depigmentation deproteinization by using a radial-flow chromatographic column which is filled with weakly basic anion exchange resin; carrying out ultrafiltration separation according to molecular weight after elution; and taking and adding an ulva fasciata delile polysaccharide compound with molecular weight of 10 KD to 30 KD into the radial-flow chromatographic column which is filled with DEAE (Diethyl Aminoethanol) sepharose gel for enriching and purifying, wherein the prepared polysaccharide with average molecular weight of 20 KD has remarkable antitumor activity.

The invention relates to a separating and purifying method for preparing white muscardine silkworm anticancer-activity polysaccharide BBPW-1, which comprises the following steps: degreasing white muscardine silkworm powder under reflux of acetone-petroleum ether and 80% ethanol to remove glycosides and alkaloids, and extracting with distilled water at 80-100 DEG C to obtain crude polysaccharide; and after removing proteins from the crude polysaccharide by a Sevag process, separating out the neutral polysaccharide component by DEAE (diethylaminoethanol) sepharose ion-exchange chromatography, carrying out propylene dextrangel S-300 chromatography, taking the first eluting peak, concentrating under reduced pressure, carrying out propylene dextrangel S-500 chromatography for further purification, and carrying out freeze-drying to obtain the white muscardine silkworm anticancer-activity polysaccharide BBPW-1. The product prepared by the method provided by the invention has uniform purity, and the molecular weight is 3.67*10<6>Da; and the product has an inhibiting action on growth of human cervical cancer cells Hela and human liver cancer cells HepG2, does not have any adverse effect on growth of normal human embryo kidney cells HEK293 and mouse macrophages RAW264.7, and can be used for developing anticancer products.

The present invention relates to a method for concentrating and/or purifying prion PrP^Sc proteins by contacting prion PrP^Sc proteins with sepharose under conditions that allow for the specific and high affinity binding of the sepharose to the prion PrP^Sc proteins and removing the unbound non-prion proteins from the sepharose, as well as the same method for removing prion PrP^Sc proteins from body fluids by contacting body fluids with sepharose under conditions that allow for the specific and high affinity binding of the sepharose to the prion PrP^Sc proteins and removing the body fluid from said sepharose. In addition, the present invention is directed to a method for separating and/or enriching prion PrP^Sc proteins from PrP^C proteins by contacting prion PrP^Sc proteins and PrP^C proteins with a ligand-modified sepharose under conditions that allow for the specific and high affinity binding of the sepharose part to the prion PrP^Sc proteins and the binding of the ligand part of the sepharose to PrP^C proteins, adding a selective release agent to the sepharose-bound proteins under conditions that allow for the release of non-prion proteins and PrP^C proteins from the ligand part of the sepharose but not for the release of the prion PrP^Sc proteins, and removing the non-prion proteins and PrP^C from the sepharose. Another aspect of the present invention concerns the use of the before-mentioned methods for concentrating, purifying and/or removing prion PrP^Sc proteins.

The invention relates to a dual PCR method for detecting theileria hirci and anaplasma. The method includes the steps of finding a gene conserved seuqnece target gene locus and designing a specific primer sequence I according to the 18S rRNA gene sequence of theileria hirci, finding a gene conserved seuqnece target gene locus and designing the specific primer sequence II according to the 16S rRNA gene sequence of anaplasma, adding the upstream primer and the downstream primer of the specific primer sequence I and the upstream primer and the downstream primer of the specific primer sequence II and PCR mixed liquid into a PCR pipe, mixing the materials and adding theileria hirci DNA and anaplasma DNA extracted from a to-be-tested blood sample to the mixture, putting the PCR pipe into a PCR amplification instrument to be circulated, obtaining a PCR product, and putting the PCR product on 2% sepharose gel to be subjected to electrophoresis. The method has the advantages of being rapid, specific, sensitive, efficient, low in cost and the like and is beneficial to clinical application.

The method is used for separating nucleic acids and other similar constructs. It involves selective introduction, enhancement, or stabilization of affinity handles such as single-strandedness in the undesired (or desired) nucleic acids as compared to the usual structure (e.g., double-strandedness) of the desired (or undesired) nucleic acids. The undesired (or desired) nucleic acids are separated from the desired (or undesired) nucleic acids due to capture by methods including but not limited to immobilized metal affinity chromatography, immobilized single-stranded DNA binding (SSB) protein, and immobilized oligonucleotides. The invention is useful to: remove contaminating genomic DNA from plasmid DNA; remove genomic DNA from plasmids, BACs, and similar constructs; selectively separate oligonucleotides and similar DNA fragments from their partner strands; purification of aptamers, (deoxy)-ribozymes and other highly structured nucleic acids; Separation of restriction fragments without using agarose gels; manufacture recombinant Taq polymerase or similar products that are sensitive to host genomic DNA contamination; and other applications

The invention relates to an affinity chromatography medium for separating and purifying a GABA receptor and a preparation method thereof. The technological process comprises the following steps: carrying out chemical modification on fipronil to obtain an affinity ligand; by taking agarose gel as a matrix, after activating by an epoxy activating agent, coupling the fipronil affinity ligand to obtain an affinity medium; packing the affinity medium so as to be used for separating and purifying fish GABA receptor protein. The affinity chromatography medium synthesized according to the invention has the advantages of being good in selectivity, strong in specificity, high in mechanical strength and good in separation effect; in addition, the preparation process is simple and the magnifying is easy.

The invention relates to a method for separating and purifying polypeptide through a hydrogen binding adsorption chromatographic medium of quercitin aglucon and high-concentration and high-crosslinking degree agarose gel. The quercitin aglucon and the high-concentration and high-crosslinking degree agarose gel serve as a medium of a matrix; and the high-purity polypeptide is further separated and purified from a biological tissue extract, a protein hydrolysate or a polypeptide synthesis mixture based on the principle of the hydrogen binding adsorption chromatography characterized by hydrogen binding mutual effect. The method has the characteristics of high selectivity, simple purification process, large sample carrying amount, high medium recycling degree and the like. The defects that the traditional method has complex steps, low reversed-phase chromatographic medium recycling degree, difficult mass production, low sample carrying amount, poor biocompatibility and the like are overcome. The purity of the obtained polypeptide fractions is determined through reversed-phase high efficiency liquid chromatography, and the sequence and the molecular weight of the polypeptide are determined through a mass spectrum.

The present invention relates to the use of milk or a derivative thereof for identifying prion proteins, preferably PrP^Sc prion proteins, in a mammal. The present invention is also directed to a method for identifying prion proteins, preferably PrP^Sc prion proteins, in mammals, comprising the step of contacting milk or a derivative thereof with an agent having high affinity and selectivity for prion proteins, preferably for PrP^Sc prion proteins. In addition, a further aspect the present invention concerns a method for removing PrP^C and/or PrP^Sc prion proteins, preferably PrP^Sc prion proteins, from milk or a milk derivative wherein milk or a derivative thereof is contacted with sepharose, preferably sepharose comprising divalent immobilized metal ions.

The invention relates to a gene engineering antibody technique and a molecular cloning technique and in particular relates to an aflatoxin B1 nano antibody immune adsorption material which is an agarose gel coupled with an aflatoxin B1 nano antibody fusion protein. The amino acid sequence of the aflatoxin B1 nano antibody fusion protein is shown in SEQ ID NO.1, and the nucleotide sequence of the protein is shown in SEQ ID NO.2. A gene engineering technique is adopted, a single-domain heavy chain antibody specifically aiming at aflatoxin B1 is fused with a self-coupled tag protein, then purification-free specific one-step orientated coupling of a recombinant expression ligand can be achieved, preparation procedures of an affine adsorption material can be greatly simplified, the cost can belowered, and meanwhile, occupation of a ligand combination site of a conventional coupling method is avoided, so that the adsorption capacity of an immune affine adsorption material can be increased.

The invention discloses a sepharose gel plate punching structure. The punching structure comprises a supporting stand, a lifting unit, and a plurality of needle tubes; the supporting stand comprises atop plate, a supporting plate and a bottom plate which are arranged in sequence from top to bottom; the bottom plate is provided with a placing surface for placing a sepharose gel plate; the liftingunit comprises at least one supporting plate lifting assembly and at least one top plate lifting assembly, the supporting plate lifting assembly drives the supporting plate to go up and down and the top plate lifting assembly drives the top plate to go up and down; negative pressure assemblies are arranged inside the needle tubes; and each of the negative pressure assemblies comprises a piston which moves up and down inside a punching tube and a piston rod with the lower end connected with the piston, the upper end of the piston rod penetrates through the punching tube and is fixed with the top plate, a through hole is formed in the upper end of the punching tube and is for the piston rod to penetrate, and the needle tube is fixed with the supporting plate. The punching structure can meetactual requirements of different experiments and rapidly punch needed holes.

The invention discloses a CAPS (Cleaved Amplified Polymorphic Sequence) marking method for a stearoyl acyl carrier protein dehydrogenase gene (SAD). The method comprises the following steps: I, extracting genome DNA (Deoxyribonucleic Acid); II, carrying out DNA sequence checking, sequence analysis and primer designing on an SAD gene; III, carrying out a PCR (Polymerase Chain Reaction) and productrecycling and purification; IV, carrying out PCR product sequencing; IV, marking CAPS, namely comparing SAD gene sequences obtained from sequencing of two varieties through BLASTN (Basic Local Alignment Search Tool), analyzing SNP (Single Nucleotide Polymorphism) sites, analyzing whether the SNP sites can be converted into CAPS markers by using software on the basis of the found SNP sites, carrying out PCR amplification on a target segment, selecting a corresponding restriction enzyme to carry out an enzyme digestion reaction, and carrying out product detection with sepharose gel, that is, analyzing CAPS marker types and domestication capabilities of parents and F1 and BCl groups by using software SPSS (Statistic Package for Social Science). The invention aims to provide beneficial assistance for molecular marker assistant selection on genotypes with relatively good cold domestication capability, and a very good effect is achieved.

The invention discloses symphytum officinale polysaccharide based on response surface method optimized enzyme-assisted extraction and a preparation method and application thereof, and belongs to the technical field of raw material deep processing technologies. The preparation method of the symphytum officinale polysaccharide comprises the following steps: step 1, extracting; step 2, alcohol precipitation and impurity removal; step 3, deproteinizing; step 4, dialyzing and desalting; step 5, carrying out DEAE-52 cellulose column chromatography; step 6, carrying out Sepharose CL-6B agarose gel column chromatography. the invention also provides an application of the symphytum officinale polysaccharide in regulating and controlling the abundance of probiotics in chicken intestinal tracts, and a complete and feasible technical route for extracting, separating and purifying the symphytum officinale polysaccharide and researching physicochemical properties and biological activity of the symphytum officinale polysaccharide is established.