1009 results about "Polyacrylamide gel electrophoresis" patented technology

A crystallizable, purified chondroitinase ABC having a molecular weight of about 100,000 dalton by the measurement of the SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and the measurement by the gel permeation chromatography method, having alanine as the N-terminal amino acid and proline as the C-terminal amino acid. A process for the purification of the crystallizable purified chondroitinase ABC comprising removing nucleic acid from an surfactant solution extract obtained from cells of chondroitinase ABC-producing microorganisms and chromatographically treating by concentration gradient elution using a weak cation exchange resin or a strong cation exchange resin. A composition comprising a chondroitinase and serum albumin, gelatin, or a nonionic surfactant.

The present invention is concerned with non-soluble proteins and soluble or insoluble fragments thereof, which bind TNF, in homogeneous form, as well as their physiologically compatible salts, especially those proteins having a molecular weight of about 55 or 75 kD (non-reducing SDS-PAGE conditions), a process for the isolation of such proteins, antibodies against such proteins, DNA sequences which code for non-soluble proteins and soluble or non-soluble fragments thereof, which bind TNF, as well as those which code for proteins comprising partly of a soluble fragment, which binds TNF, and partly of all domains except the first of the constant region of the heavy chain of human immunoglobulins and the recombinant proteins coded thereby as well as a process for their manufacture using transformed pro- and eukaryotic host cells.

A protein from M. catarrhalis, designated the 74 kD protein, is isolated and purified. The 74 kD protein has an amino-terminal amino acid sequence which is conserved among various strains of M. catarrhalis. The protein has a molecular weight of approximately 74.9 kD as measured on a 10% SDS-PAGE gel, while its molecular weight as measured by mass spectrometry is approximately 74 kD. The 74 kD protein is used to prepare a vaccine composition which elicits a protective immune response in a mammalian host, to protect the host against disease caused by M. catarrhalis.

The invention provides diacylglycerol acyltransferase (DAGAT) proteins, wherein said proteins are active in the formation of triacylglycerol from fatty acyl and diacylglycerol substrates. In one aspect, Mortierella ramanniana DAGAT proteins have been isolated and have molecular weights of between approximately 36 and 37 kDa as measured by SDS-PAGE. The invention also provides novel DAGAT polynucleotide and polypeptide sequences and to methods of producing such polypeptides using recombinannt techniques. In addition, methods are provided for using such sequences to alter triacylglycerol levels in plants and to treat diseases associated with altered DAGAT activity or expression.

There are provided a soluble CD14 antigen which is a novel in vivo protein useful as a marker for diagnosing sepsis and has the following characteristic features 1) to 3):

• • 1) a molecular weight of 13±2 kDa when measured by SDS-PAGE under non-reducing conditions;
  • 2) an amino acid sequence in which the amino acid sequence of SEQ ID NO:1 is present on its N terminal; and
  • 3) ability to specifically bind to an antibody prepared by using a peptide comprising 16 amino acid residues described in SEQ ID NO:2 for the antigen; and a recombinant soluble CD14 fragment.

There are provided natural rubber free from substances which cause Type I allergy, a rubber compositions having good processability and physical properties which comprises the natural rubber and other rubber, and a tire product comprising the natural rubber. The natural rubber of the present invention exhibits the above properties by containing substantially no proteins specified by the bands of 14, 31 and 45 kDa by SDS-PAGE.

The invention discloses an anti-rabies virus monoclonal antibody and a preparation method and an application, belonging to the field of biomedicine and particularly relating to the preparation of a monoclonal antibody capable of identifying rabies virus and the application. The monoclonal antibody of the invention is screened by indirect Enzyme-linked immunosorbent assay (ELISA), and specificity and affinity thereof combined with antigen are identified by methods such as polyacrylamide gel electrophoresis analysis, speckle ELISA, immunoblot analysis and the like. The anti-rabies virus monoclonal antibody of the invention can be applied in multiple testing methods of antigen of rabies virus and can be also applied in the preparation of rabies virus detecting kit. The anti-rabies virus monoclonal antibody is secreted by anti-rabies virus monoclonal antibody hybridoma cell strain 2C5 with the preservation number being CGMCC No.3014.

A water soluble extract of M.charantia named MC6, methods for its preparation and methods for its use in the treatment of hyperglycemic disorders are provided. The active MC6 is characterized by moving as a single band on SDS-PAGE having a molecular weight of less than 10 kDal, and by comprising three peptides. Also provided is a peptide component of MC6 named MC6.1, as well as analogues and mimetics thereof. The active MC6, MC 6.1, MC6.2, and MC6.3 exhibit hypoglycemic activity, even following oral administration. Also provided are methods of using the active agents to treat hyperglycemic disorders, particularly diabetes, where the active agents are preferably orally administered.

The present invention relates to a method for detecting immunoblotting based on quantum dot labeling. Firstly, connecting quantum dot with carboxyl or amido with second antibody by chemical reaction to form compound with quantum dot label; employing polyacrylamide gel electrophoresis, analytes is protein, probe is antibody, quantum dot labeled second antibody is used to show color. Protein sample separated by PAGE is transferred to solid carrier, the protein or polypeptide on solid carrier as antigen is processed immunoreaction with corresponding antibody, and then reacted with quantum dot labeled second antibody, under the UV lamp, quantum dot emitting fluorescence to detect electrophoresis separated specific objective gene expression protein component. Immunoblotting image is gained by filter equipped of simple UV imaging system. The present invention employs quantum dot connected with second antibody as excellent fluorescence imaging agent of immunoblotting to enhance the sensitivity and precision.

The invention discloses a recombinant antigenic protein for diagnosing echinococcosis granulosus (having an amino acid sequence represented by SEQIDNo.1). In addition, the invention also discloses the preparation method of the recombinant antigenic protein, which comprises: amplifying an EgEPC1 gene by using RT-PCR; cloning the EgEPC1 gene in a pGEM-T vector; connecting the EgEPC1 gene with an expression vector PET28a(+) to form a recombinant plasmid PET28a-EgEPC1; transforming the recombinant plasmid PET28a-EgEPC1 to Escherichia coli BL21(DE3) and expressing the recombinant protein through IPTG induction; and identifying a purified recombinant antigen by using SDS-PAGE and Western blotting. In addition, the invention also discloses the diagnosis use of the recombinant antigenic protein. Experiments show that the recombinant antigenic protein of the invention has the advantages of high sensibility and specificity for the diagnosis of echinococcosis granulosus, and has a promising application prospect in the diagnosis of the echinococcosis granulosus.

The invention provides a construction method for a DNA fingerprint database of a high flux cotton variety. The method comprises the following steps: extracting DNAs of the cotton variety, carrying out SSR-PCR amplification by using 40 pairs of primers (with nucleotide sequences as represented by SEQ ID No. 1-80), carrying out fluorescence multiple PCR combination on products obtained after amplification, carrying out capillary five-color fluorescence electrophoresis detection with a DNA analyzer and constructing the DNA fingerprint database of the cotton variety according to a fluorescence spectrum. Compared to detection efficiency of conventional polyacrylamide gel electrophoresis, detection efficiency of the method provided in the invention is increased by more than 10 times; in addition, since the size of fragments of the amplification products is obtained through automatic comparison of internal standard of molecular weight, human errors in plate reading is reduced, and accuracy and reliability of data results are utmostly guaranteed at the same time.

A protein which inhibits osteoclast differentiation and/or maturation and a method of production of the protein. The protein is produced by human embryonic lung fibroblasts and has molecular weight of about 60 kD and about 120 kD under non-reducing conditions and about 60 kD under reducing conditions on SDS-polyacrylamide gel electrophoresis, respectively. The protein can be isolated and purified from culture medium of the said fibroblasts. Furthermore, the protein can be produced by gene engineering. The present invention includes cDNA for producing the protein by gene engineering, antibodies having specific affinity to the protein or a method for determination of the protein concentration using the antibodies.

The invention provides a method for establishing an SSR (Simple Sequence Repeat) fingerprint spectrum of broad beans, and belongs to the technical field of plant variety identification and breeding. The method provided by the invention provides a marker combination consisting of 21 pairs of broad bean genome SSR primers, and the nucleotide sequences of which are shown in SEQ ID NO. 1 to 42 respectively; the broad bean varieties are identified by PCR (Polymerase Chain Reaction) amplification technology, non-denaturing polyacrylamide gel electrophoresis or capillary electrophoresis detection technology, and the identification of broad bean seed varieties and genetic diversity assessment work can be completed in a short time. The method has the advantages of time saving, high efficiency, fastness, accuracy, and convenience in operation, and the identification results are not easily affected by the environment. The method provided by the invention has the advantages of effectively monitoring the authenticity of the broad bean seeds, revealing the genetic variation and genetic relationship of the varieties from the DNA level, protecting the crop varieties, preventing the fake and inferior varieties from entering the market, and also providing a technical support for the reasonable utilization of excellent germplasm during the breeding of broad bean varieties, and has good application prospects.

The present invention discloses a cabbage SSR fingerprint construction method, and belongs to the technical field of agricultural cabbage breeding and application. DNA of a variety to be tested is extracted. Twenty nine pairs of core SSR primers as shown in a sequence table SEQ ID No. 1-58 are respectively used for PCR amplification of the DNA of the variety to be tested. PCR products are detected by a denatured polyacrylamide gel electrophoresis or a five-color fluorescent capillary electrophoresis to construct cabbage variety SSR fingerprints. The invention establishes a fast and accurate, economic and simple, stable and reliable cabbage DNA fingerprint certification standard experimental system, and on this basis, develops a DNA fingerprint detection method applicable for cabbage variety identification. The present invention utilizes capillary electrophoresis methods for variety identification of cabbage, enhances the experimental efficiency in time, and has the advantages of fast speed, accuracy and easy operation, and identification results are not influenced by environments.

The present invention uses an optimized albumen extracting method, can test the constituent and the content of the foam albumen in the beer barley and the malt using the dodecyl sodium sulfate - polyacrylamide gel (SDS-PAGE) electrophoresis technology, and can detect the change of the constituents and the content of the foam albumen during the germinating process of the beer barley and the malt.

The invention discloses a method for constructing a wheat SSR fingerprint, which belongs to the field of agricultural wheat breeding and application technologies. The method comprises the following steps: extracting DNA (deoxyribonucleic acid) of a wheat variety to be tested; carrying out PCR (polymerase chain reaction) amplification to the DNA Of the wheat variety to be tested by using 13 pairs of basic core SSR fluorescent dye primers with the ID No.1 to 26 and shown in a sequence table SEQ and 12 pairs of extended core SSR fluorescent dye primers with the ID No.27 to 50 and shown in the sequence table SEQ respectively; and detecting PCR amplification products by denaturing polyacrylamide gel electrophoresis or five-color fluorescent capillary electrophoresis, and constructing the SSR fingerprint of the wheat variety. The method provided by the invention can identify the wheat variety in a laboratory by using technologies of the PCR amplification, the gel electrophoresis and capillary electrophoresis according to the polymorphism of SSR sites, can greatly improve the test efficiency in time, and has the advantages of high speed, high accuracy, convenience for operation and the like; and the identification result is not liable to be affected by the environment.