Abstract: Novel compounds comprising a C—C double bond substituted at one carbon with two sulfur atom-containing groups are disclosed. The compounds are useful in methods and compositions for generating chemiluminescence rapidly by reaction with a peroxidase enzyme and a peroxide. The chemiluminescence thus produced can be used as a detectable signal in assays for peroxidase enzymes or peroxide-producing enzymes and in assays employing enzyme-labeled specific binding pairs.

Abstract: Methods are disclosed for producing electrochemiluminescence by electrochemically oxidizing an acridan compound at an electrode in the presence of a peroxide. Maintaining a sufficiently positive potential results in continuous oxidation of the acridan compound to an acridinium compound which reacts with peroxide to produce the luminescence. Light emission can be reversibly and repeatedly cycled on and off by sweeping the potential between two values. The acridan compounds can be provided with a labeling group for linking to an analyte or analyte binding partner. The present electrochemiluminescent reaction can find use in assay methods for detecting analytes by immunoassays and nucleic acid assays.

Abstract: Methods of detecting or measuring the activity of polynucleotide kinase are disclosed as well as methods of detecting an analyte in an assay using polynucleotide kinase as a label on a member of a specific binding pair. the methods rely on the phosphorylation of an oligonucleotide followed by ligation of the oligonucleotide 5?-phosphate onto another template-bound oligonucleotide. The presence of the ligated product signals the presence of polynucleotide kinase. In preferred embodiments, phosphorylation of an oligonucleotide enables the consecutive ligation of a set of oligonucleotides. The oligonucleotides so ligated can be detectably labeled with, for example, other enzymes to provide highly sensitive detection methods.

Abstract: Methods of detecting or measuring the activity of polynucleotide kinase are disclosed as well as methods of detecting an analyte in an assay using polynucleotide kinase as a label on a member of a specific binding pair. The methods rely on the phosphorylation of an oligonucleotide followed by ligation of the oligonucleotide 5?-phosphate onto another template-bound oligonucleotide. The presence of the ligated product signals the presence of polynucleotide kinase. In preferred embodiments, phosphorylation of an oligonucleotide enables the consecutive ligation of a set of oligonucleotides. The oligonucleotides so ligated can be detectably labeled with, for example, other enzymes to provide highly sensitive detection methods.

Abstract: A method of enhancing the isolation of ribonucleic acids from samples of biological or cellular material is disclosed which uses a solution of an Agent during binding of nucleic acids onto a solid phase binding material which, after elution, enhances the recovery of RNA. Washing the solid phase and eluting the nucleic acid produces RNA in enhanced yield and/or purity. The use of the new method allows RNA to be captured and released in a form suitable for downstream processing in under five minutes. Preferred Agents according to the invention comprise monomeric, oligomeric, dendrimeric and polymeric organic compounds having multiple positive charges.

Abstract: Methods using chemiluminescent label compounds and chemiluminescent labeled conjugates are provided. The compounds comprise an acridan ring bearing an exocyclic ketene dithioacetal group and further contain a labeling substituent which permits attachment to compounds of interest. The novel chemiluminescent compounds and labeled conjugates are convenient to prepare, are highly stable, and generate chemiluminescence rapidly on demand. The compounds and conjugates are useful in assays of an analyte in a sample and in assays employing labeled specific binding pairs.

Abstract: Methods for generating chemiluminescence rapidly by reaction of at least one compound comprising a C—C double bond substituted at one carbon with two sulfur atom-containing groups with a peroxidase enzyme and a peroxide are disclosed. The chemiluminescence thus produced can be used as a detectable signal in assays for peroxidase enzymes or peroxide-producing enzymes and in assays employing enzyme-labeled specific binding pairs. Use of the methods provides rapid signal generation, achieving a plateau intensity in under one minute. The substrate can be provided in a composition which demonstrates unexpectedly long storage stability.

Abstract: Compounds useful for detecting a source of hydrogen peroxide are disclosed wherein a signalling compound of the formula: is reacted with peroxide. Sig is an aromatic or heteroaromatic ring group, B is a boron atom, and R5 and R6 are independently selected from hydrogen and lower alkyl groups and can be joined together as a straight or branched alkylene chain forming a five or six-membered ring. A detectable product compound of the formula Sig-OH Is produced and detected by measuring color, fluorescence, chemiluminescence, or bioluminescence. The signalling compound itself does not possess the detectable property or does so only to a very weak degree. The compounds can be used for detection in assays for peroxide or peroxide-producing enzymes and in assays employing enzyme-labeled specific binding pairs.

Abstract: Methods of isolating nucleic acids from samples of biological or cellular material are disclosed which use solid phase binding materials and which avoid the use of any lysis solution or coating. The use of the solid phase binding materials unexpectedly allow the nucleic acid content of cells to be freed and captured directly and in one step. The new methods represent a significant simplification over existing methods. Nucleic acids can be captured and released in a form suitable for downstream processing in under five minutes. Preferred solid phase materials for use with the methods and compositions of the invention comprise a quaternary onium nucleic acid binding portion.

Abstract: Methods of producing fluorescence from fluorogenic substrates reactive with a peroxidase enzyme are disclosed. Use of the methods in assays for peroxidase enzymes, peroxidase-labeled analytes are provided. Fluorogenic compounds, compositions and kits for reaction with peroxidase enzymes are described. Two modes of producing fluorescent compounds are described.

Abstract: Methods of isolating nucleic acids from samples of cellular material are disclosed which use solid phase binding materials and which avoid the use of a lysis solution. The use of the solid phase binding materials unexpectedly allow the nucleic acid content of cells to be freed and captured directly and in one step. The new methods represent a significant simplification over existing methods. Preferred solid phase materials for use with the methods and compositions of the invention comprise a quaternary onium nucleic acid binding portion.

Abstract: Methods of isolating nucleic acids are disclosed comprising binding the nucleic acid to solid phase binding materials and releasing the bound nucleic acid from the solid phase by elution with a novel reagent composition. Compositions feature a high ionic strength buffer or an added hydrophilic organic co-solvent or both. Preferred solid phase materials for use with the methods and compositions of the invention comprise a quaternary onium nucleic acid binding portion.

Abstract: Solid phase materials for binding nucleic acids and methods of their use are disclosed. The materials feature a cleavable linker portion which can be cleaved to release bound nucleic acids. The solid phase materials comprise a solid support portion comprising a matrix selected from silica, glass, insoluble synthetic polymers, and insoluble polysaccharides to which is attached a nucleic acid binding portion for attracting and binding nucleic acids, the nucleic acid binding portion (NAB) being linked by a cleavable linker portion to the solid support portion. Preferred nucleic acid binding portions comprise a ternary or quaternary onium group. The materials can be in the form of microparticles, fibers, beads, membranes, test tubes or microwells and can further comprise a magnetic core portion. Methods of binding nucleic acids using the cleavable solid supports are disclosed as are their use in methods of isolating or purifying nucleic acids.

Abstract: Solid phase materials for binding nucleic acids and methods of their use are disclosed. The materials feature a cleavable linker portion which can be cleaved to release bound nucleic acids. The solid phase materials comprise a solid support portion comprising a matrix selected from silica, glass, insoluble synthetic polymers, and insoluble polysaccharides to which is attached a nucleic acid binding portion for attracting and binding nucleic acids, the nucleic acid binding portion (NAB) being linked by a cleavable linker portion to the solid support portion. Preferred nucleic acid binding portions comprise a ternary or quaternary onium group. The materials can be in the form of microparticles, fibers, beads, membranes, test tubes or microwells and can further comprise a magnetic core portion. Methods of binding nucleic acids using the cleavable solid supports are disclosed as are their use in methods of isolating or purifying nucleic acids.

Abstract: Novel compounds comprising a C—C double bond substituted at one carbon with two oxygen or sulfur atom-containing groups are disclosed. The compounds are useful in methods and compositions for generating chemiluminescence by reaction with a peroxidase enzyme and a peroxide. The chemiluminescence thus produced can be used as a detectable signal in assays for peroxidase enzymes and in assays employing enzyme-labeled specific binding pairs.

Abstract: Novel compounds comprising a C—C double bond substituted at one carbon with two sulfur atom-containing groups are disclosed. The compounds are useful in methods and compositions for generating chemiluminescence rapidly by reaction with a peroxidase enzyme and a peroxide. The chemiluminescence thus produced can be used as a detectable signal in assays for peroxidase enzymes or peroxide-producing enzymes and in assays employing enzyme-labeled specific binding pairs.

Abstract: Novel compounds comprising a C—C double bond substituted at one carbon with two sulfur atom-containing groups are disclosed. The compounds are useful in methods and compositions for generating chemiluminescence rapidly by reaction with a peroxidase enzyme and a peroxide. The chemiluminescence thus produced can be used as a detectable signal in assays for peroxidase enzymes or peroxide-producing enzymes and in assays employing enzyme-labeled specific binding pairs.

Abstract: Methods of detecting or measuring the activity of polynucleotide kinase are disclosed as well as methods of detecting an analyte in an assay using polynucleotide kinase as a label on a member of a specific binding pair. The methods rely on the phosphorylation of an oligonucleotide followed by ligation of the oligonucleotide 5′-phosphate onto another template-bound oligonucleotide. The presence of the ligated product signals the presence of polynucleotide kinase. In preferred embodiments, phosphorylation of an oligonucleotide enables the consecutive ligation of a set of oligonucleotides. The oligonucleotides so ligated can be detectably labeled with, for example, other enzymes to provide highly sensitive detection methods.

Abstract: Novel compounds comprising a C—C double bond substituted at one carbon with two sulfur atom-containing groups are disclosed. The compounds are useful in methods and compositions for generating chemiluminescence rapidly by reaction with a peroxidase enzyme and a peroxide. The chemiluminescence thus produced can be used as a detectable signal in assays for peroxidase enzymes or peroxide-producing enzymes and in assays employing enzyme-labeled specific binding pairs.

Abstract: Compounds useful for detecting a source of hydrogen peroxide are disclosed wherein a signalling compound of the formula: 1