266 results about "Immunoprecipitation" patented technology

The present invention provides systems, devices, apparatuses and methods for automated bioprocessing. Examples of protocols and bioprocessing procedures suitable for the present invention include but are not limited to: immunoprecipitation, chromatin immunoprecipitation, recombinant protein isolation, nucleic acid separation and isolation, protein labeling, separation and isolation, cell separation and isolation, food safety analysis and automatic bead based separation. In some embodiments, the invention provides automated systems, automated devices, automated cartridges and automated methods of western blot processing. Other embodiments include automated systems, automated devices, automated cartridges and automated methods for separation, preparation and purification of nucleic acids, such as DNA or RNA or fragments thereof, including plasmid DNA, genomic DNA, bacterial DNA, viral DNA and any other DNA, and for automated systems, automated devices, automated cartridges and automated methods for processing, separation and purification of proteins, peptides and the like.

The invention provides a genetic screening method for identifying a transfected cell expressing the polypeptide of interest. The methods allows for high throughput screening of recombinant cells for elevated levels of expression of the polypeptide of interest. The invention also provides capture media, formulations and methods of making and using thereof.

The invention provides a method of mapping DNA-protein interactions within a genome by fixing living cells to cross-link DNA and proteins, lysing the cells, and isolating chromatin by immunoprecipitation. DNA is purified and a SAGE protocol is performed on the purified DNA to produce GMAT-tag sequences, which are compared to a genomic sequence of the living cells to map DNA-protein interactions. The invention further provides a method of identifying an active chromatin domain and a method of identifying aberrant chromatin acetylation, wherein chromatin immunoprecipitation is performed using an antibody recognizing acetylated histone protein.

In various aspects, the present invention provides methods and kits for predicting the therapeutic efficacy of an immunological reagent, identifying an immunological reagent having therapeutic efficacy, or both, for the treatment of an amyloidogenic disorder by comparing the amount of Aβ monomer in an Aβ preparation which binds to the immunological reagent to an amount of one or more Aβ oligomers in the Aβ preparation which bind to the immunological reagent to determine a relative bound amount, and predicting the efficacy of the immunological reagent, identifying an immunological reagent having therapeutic efficacy, or both, for the treatment of an amyloidogenic disorder based at least on the relative bound amount.

Antibodies are described, which are suitable for identification of gamma-carboxyglutamic acid (Gla), said antibodies having an ability of identifying Gla residues in proteins and/or peptides, while not reacting with glutamic (Glu) residues in corresponding proteins and/or peptides. Methods for the preparation and identification of said antibodies as well as methods for detection of Gla in biological fluids, tissue extracts, tissue specimens, in which methods said antibodies are used, are also described. There is also described the use of said antibodies for immunopurification of Gla-containing proteins or peptides by, for instance, immunoprecipitation or immunoaffinity chromatography.

The invention provides methods and compositions for noninvasive prenatal diagnosis of fetal aneuploidies. A large panel of differentially methylated regions (DMRs) have been identified. Certain of these DMRs are hypomethylated in adult female blood DNA and hypermethylated in fetal DNA, whereas others are hypermethylated in adult female blood DNA and hypomethylated in fetal DNA. Moreover, DMRs that are hypomethylated in adult female blood DNA and hypermethylated in fetal DNA have been shown to accurately predict a fetal aneuploidy in fetal DNA present in a maternal blood sample during pregnancy. In the methods of the invention, hypermethylated DNA is physically separated from hypomethylated DNA, preferably by methylated DNA immunoprecipitation.

The present invention provides a high-throughput sequencing method for methylated DNA, and use thereof. Particularly, the present invention provides a high-throughput sequencing method for methylated DNA, which combines methylated DNA immunoprecipitation, removal of repetitive sequences, and bisulfite treatment. The site of sequencing library will be decreased, and the cost will be reduced by using the method disclosed in the present invention.

The following invention outlines methodologies for the construction and utilization of transcription factor direct target gene microarrays of both DNA and corresponding protein/peptide target origin. The technology entails the array/microarray annotation and organization of transcription factor direct loci and corresponding protein products identified through modified and improved versions of chromosomal immunoprecipitation (CHIP) and molecular cloning procedures. It allows for the formulation of physiologically directed arrays which result in a thorough, focused characterization of the genetic and biochemical regulation occurring within a give population of cells or a given tissue. Arrays and microarrays of direct targets for any given transcription factor created utilizing this technology are substantially more clinically relevant for purposes of medical diagnostics and patient prognostics than conventional microarrays due to the physiologically focused nature and the transcription factor targets. In addition, the characterization and array organization of transcription factor target protein products and the assessment of their interactions with other proteins and/or small molecules is of critical importance for the purposes of understanding cellular and ultimately the design of therapeutics for human anomalies.

The invention discloses a lyophilized viper antivenin and a preparation method thereof, belonging to biochemical products, more particularly relating to an antivenin and a preparation technology thereof. The mass ratio of the antivenin and viper venom is 15:1, which can specifically neutralize the viper venom injected to mouse; as determined by immunodiffusion, the immunoprecipitation line appears in case that the ratio of viper antivenin F(ab')2 in lyophilized form to viper venom is 8:1; and other detected items conform to the quality standard of antivenin in Chinese Pharmacopoeia 2010. The detection of Phenyl-Sepharose (low-sub) FF column chromatography result shows that: activity is centralized at eluting peak 1, micromolecule impurity proteins are centralized at penetration peal, eluting peak 2 and eluting peak 3. According to the technology of the invention, immune blood plasma is resulted from viper venom immune horse, IgG is prepared by salting out the immune blood plasma, and F(ab')2 active fragment is obtained after the IgG is subject to enzymolysis and purification by a hydrophobic column. The lyophilized viper antivenin has strong specificity, high potency and more than 85% of the purity of antivenin F(ab')2 in lyophilized form.

An ELISA kit for detecting a Salmonella pullorum antibody is established by screening Salmonella pullorum dominant antigen GroEL through an immunoprecipitation technology, expressing GroEL recombinant protein through utilizing a prokaryotic expression vector, and utilizing an antigen protein. The kit can reduce the response of chicken in the detection process of the Salmonella pullorum antibody, and can improve the detection specificity and the sensitivity.

The invention discloses an anti-human CEACAM5 monoclonal antibody and a preparation method and application thereof, and the anti-human CEACAM5 monoclonal antibody is produced by a mouse hybridoma cellline with the accession number CCTCC NO: C2016129. The preparation method comprises the following steps: S1, immunizing a mouse with whole cell protein, lysed by human gastric cancer cells, as an immunogen; S2, fusing spleen cells of the immunized mouse obtained in the step S1 with mouse myeloma cells; S3, selecting a hybridoma cell line stably secreting antibodies after cell cloning occurs during the cell fusing in the step 2; and S4, preparing the anti-human CEACAM5 monoclonal antibody from the hybridoma cell line stably secreting the antibodies obtained in the step 3. The monoclonal antibody can be used to prepare a composition for treating a malignant tumor. The anti-human CEACAM5 monoclonal antibody can be widely used in various routine operations such as flow cytometry, immunohistochemical staining and immunoprecipitation.

The present disclosure is directed to antibody-linked immuno-sedimentation agent, the antibody being linked to a sedimentation agent by a non-antigen binding region of the antibody, and a method of isolating a target from a sample using the antibody-linked immuno-sedimentation agent. The methods involve forming a mixture including a sample with an antibody linked immuno-sedimentation agent and red blood cells under conditions sufficient to form red blood cell rouleaux and allow antibody-antigen binding.

The present invention relates to a method for detecting antibodies against a target antigen in a sample which comprises contacting the sample with labelled target antigen, subjecting the sample to immunoprecipitation to precipitate antibodies in the sample and detecting the presence of antibodies against the target antigen in the sample by means of the presence of labelled target antigen in the immunoprecipitate, wherein the labelled target antigen is a fusion protein comprising the target antigen and a fluorescent protein label and the presence of labelled target antigen in the immunoprecipitate is detected by means of the fluorescence of the fluorescent label. The method is particularly suitable for use where the target antigen is an autoantigen and can also be used to identify autoantigens implicated in a particular autoimmune disorder by screening serum samples from patients with a clinical phenotype indicative or suggestive of an autoimmune disorder and suitable controls. The target protein may be from the cys-loop acetyl choline receptor ion channel gene superfamily, the voltage-gated calcium, sodium or potassium ion channel gene superfamily, the glutamate receptor gene family, a receptor tyrosine kinase, or other membrane associated channels such as aquaporin gene family.

The invention relates to the field of plant antiviral genetic engineering, and provides a plant antiviral new target. On the basis of tobacco vein banding mosaic virus infection cloning, photosystem II oxygen-evolving complex protein PsbO1 is screened through immunoprecipitation and mass spectrography, and a PbsO1 gene is obtained through reverse transcription PCR amplification. Immunoprecipitation and bimolecular fluorescence complementation prove that the PsbO1 can interact with the tobacco vein banding mosaic virus 6K2. Reduction in expression of the PsbO1 gene enables an infected plant to generate resistance to the tobacco vein banding mosaic virus and potato virus Y.

The present invention describes methods of producing milligram quantities of three forms of purified Stat1 protein from recombinant DNA constructs. In addition, the Stat proteins may be isolated in their phosphorylated or nonphosphorylated forms (Tyr 701). The proteins can be produced in baculovirus infected insect cells, or E. coli. A compact domain in the amino terminus of Stat1α was isolated and found to enhance DNA binding due to its ability to interact with a neighboring Stat protein. A relatively protease-resistant recombinant truncated form of the Stat protein was isolated in 40-50 mg quantities. Purification of the Stat proteins were performed after modifying specific cysteine residues of the Stat proteins to prevent aggregation. Activated EGF-receptor partially purified from membranes by immunoprecipitation was shown to be capable of in vitro catalysis of the phosphorylation of the tyrosine residue of Stat1 known to be phosphorylated in vivo. Techniques are enclosed to separate the phosphorylated from the nonphosphorylated Stat proteins. The techniques disclosed are general for Stat proteins and may be used to isolate large quantities of purified Stat 2, 3, 4, 5A, 5B and 6. Methods for using purified Stat proteins, truncated Stat proteins, or Stat N-terminal fragments for drug discovery are also disclosed.

The invention provides a method and kit for detecting RNA N6-methyladenosine modification at single-base resolution in a range of a whole transcriptome. According to the method, on the basis of an N6-allyl label of in-vivo ribonucleic acid (RNA) adenine and chemical treatment, base mutation of the in-vivo ribonucleic acid (RNA) adenine in a process of reverse transcription into DNA is induced, andthen a mutation site is recognized by means of nucleic acid sequencing, so that an a6A site is obtained, and the a6A site is a site originally modified by m6A in cell RNA. By means of the method, thespecific label of N6-allyladenine in a cell is achieved for the first time, and the label not only can be used for replacing an N6-methyladenosine site in the cell, but also can be positioned by means of mutation sequencing. Compared with existing gene sequencing technologies applied to m6A detection, the method for detecting RNA N6-methyladenosine modification at single-base resolution in the range of the whole transcriptome has the advantage that due to the fact that the mutation site can be accurate down to single-base resolution, the precision of m6A site detection based on m6A antibody immunoprecipitation and a massively parallel sequencing method which are currently and generally adopted is improved, so that the method for detecting RNA N6-methyladenosine modification at single-baseresolution in the range of the whole transcriptome is a direct high-throughput single-base identification method.

The present invention is based on the discovery that an antibody specific for PDH complex can be used to immunoprecitate PDH complex from the patient sample in an active state. Therefore the anti-PDH complex specific antibody can be used to determine the amount of and/or active state of PDH in a patient sample. The invention immunoassay methods for determining the amount and/or active state of PDH complex present in a patient sample are useful for screening to identify individuals having symptoms indicating malfunction of PDH complex. For example, the invention methods can be used to screen individuals for symptoms of onset of the diabetic state, such as insulitis, and for diagnosing late onset diseases, such as diabetes, Alzheimer's and the like.

A device termed the MobileArray™ device and methods, which form the MobileArray™ system, are disclosed for performing multiplex assays, target enrichment or purification. The device and methods disclosed enable the performing of multiplex assays, target enrichment or purification in a simplified manner. The MobileArray™ device gains the advantages and applications of the advanced multiplexing platforms, but does not require the special expensive equipment, reagents, software or dedicated operators. In addition, the MobileArray™ system can also be utilized in immunoprecipitation and target enrichment or purification in a multiplex manner. Furthermore, the MobileArray™ system can be integrated in an automated procedure. The MobileArray™ system makes it possible to apply multiplexing protocols in routine clinical practice, food safety inspection and general life science research laboratories.

The invention relates to a phosphorylation antibody chip kit for detecting human RTK. The phosphorylation antibody chip kit for detecting human RTK comprises: a solid-phase vector, which is a standard film or slide coated with a specific antibody; a cleaning solution, which comprises a 20X concentrated cleaning solution of 0.1% Tween-20 and a dilution solution thereof; a sample diluting solution; a dilution solution used for diluting the antibody and HRP-streptavidin; a biotinylated detection antibody mixture; a 300X concentrated fluorescein-streptavidin solution; a sample treating solution, which is a 2X cell lysis buffer; and the specific antibody, which is targeted to 71 proteins. The RTK phosphorylation antibody chip kit provided by the invention can rapidly, simply and accurately determine the situations of phosphorylation of 71 proteins in a RTK pathway through one experiment; and pathway activation in cells can be rapidly detected by monitoring the phosphorylation changes of proteins in an experimental model system without complicated immunoprecipitation and blotting.