Abstract: Methods for internally and externally photostandardizing chemical assays, e.g. an ATP-bioluminescence assay. A pre-determined amount of a photosensitive derivative of the analyte of interest is used in the assay protocol which releases a known amount of the free analyte when it is exposed to a flash of visible light of pre-determined duration and intensity. By monitoring the response of a test property of the assay to the release of a known quantity of analyte, a standard value can be calculated which allows either direct determination of the amount of analyte originally present in the assayed sample or production of a photocalibration series against which results of an assayed sample can be compared.

Abstract: Methods for identifying inducers and inhibitors of programmed cell death in a cell-free system are described. The methods exploit the finding that programmed cell death is accompanied by shutdown of cellular protein synthesis and by phosphorylation of eIF-2.alpha. and that the dephosphorylation of eIF-2.alpha. prevents the shutdown of protein synthesis.

Abstract: A sample is treated with a phosphoglyceric acid mutase (PGAM) inhibitor (polythionic acid (salt) such as potassium tetrathionate) to inactivate the M-type isozyme activity and the B-type PGAM is quantified by determining PGAM isozymes by a rate assay. The B-type PGAM is a novel marker for cerebral apoplexy, and the diagnosis of cerebral apoplexy is enabled by assaying it.

Abstract: The present invention discloses a bacteria-coding identification method,bacteria biochemical properties identifying papers which detect the biological properties using this method, and apparatus or means useful for identifying the genus and species of bacteria using the coding identification method and biochemical properties-identifying papers.

Abstract: The invention relates to isolated nucleic acids encoding recombinant calf intestinal alkaline phosphatase. Expression vectors and host cells transformed or transfected with such vectors are also provided. The invention further provides multifunctional polypeptides containing amino acid sequences encoding for calf intestinal alkaline phosphatase and a second amino acid sequence encoding a reagent having specific reactivity with a ligand. The recombinant calf intestinal alkaline phosphatase or its active fragments and the multifunctional polypeptides can be used in the methods for determining the presence or concentration of a ligand.

Abstract: The subject invention provides for nucleotide sequences encoding polypeptide p62 and derivatives thereof. Another aspect of the subject invention also provides for methods of purifying p62 and derivatives thereof from cells naturally producing p62 and from cells genetically modified so as to produce p62.The subject invention also provides for methods of assaying tyrosine kinase activity by means of measuring the phosphorylation of p62 and p62 derivatives. Measurement of p62/p62 derivative phosphorylation may be used to determine whether or not a call is cancerous.

Abstract: The subject invention pertains to a combination of enzyme activities comprising an ATP-degrading enzyme and one or more enzymes capable of degrading substances, other than ATP, that are substrates for the light-emitting reaction of firefly luciferase. These activities can be used to treat a growth medium in order to reduce background to negligible amounts, in a subsequent bioluminescence assay.

Abstract: The invention is directed to rapid and quantitative assay systems for screening test compounds for their ability to modulate tyrosine kinase or phosphatase activities involved in signal transduction by determining the tyrosine phosphorylation state of a protein substrate using an anti-phosphotyrosine antibody and an antibody specific for the protein substrate. These assays may be practiced in a whole cell or cell-free system. The assays can be used to identify test compounds for use in therapeutic applications to disease processes in which tyrosine kinase or phosphatase activity in a signal transduction pathway contributes to a pathological process.

Abstract: A method for simultaneously testing a sample for the presence of multiple rget antigens or antibodies in the sample, which comprises presenting the sample to a plurality of different binding sites, wherein at least two of the sites are binding sites for different known target antigens or antibodies, each known binding site is composed of at least one molecule of a ligand-enzyme complex attached to a support, and the ligand-enzyme complex is a ligand attached to an enzyme in proximity to the enzyme's active site such that the enzymatic activity of the ligand-enzyme complex is changed when the target antibody or antigen is present in the sample. The ligand-enzyme complex embraces nonenzymatic reporter molecules such as electrochemiluminescent compounds. The method also includes assaying each binding site for a change in enzymatic or other reporter activity compared to a control value. A device for performing simultaneous detection of multiple target antigens or antibodies is also disclosed.

Abstract: This invention provides materials and methods for identifying inhibitors of a Hepatitis C Virus NS3 protein ATPase. Methods for making and purifying such an ATPase are also provided by this invention.

Abstract: The present invention provides a culture medium for microbiological tests prepared by treating medium components containing adenosine triphosphate (ATP) with an acidic phosphatase as well as a microbiological test method characterized by using said culture medium for microbiological tests as the 1st aspect of the present invention when the microbiological test is conducted by ATP-luciferase method. When using the culture medium for microbiological tests according to the present invention, in a microbiological test by ATP-luciferase method, the emission derived from the culture medium is suppressed stably at a very low level, and there is no possibility that the emission derived from a substance other than a microorganism affects the measurement. In addition, the microbiological test method according to the present invention is simple, accurate and highly reliable since it employs the culture medium for microbiological tests according to the present invention.

Abstract: A chemiluminescent assays for the determination of the presence or amount of a biopolymer in bound assays using 1,2-dioxetanes in connection with AttoPhos.TM. as chemiluminescent substrates for enzyme-labeled targets or probes is provided. Further disclosed is a kit for conducting a bioassay for the presence or concentration of a biopolymer comprising a) an enzyme complex; b) a 1,2-dioxetane; and c) AttoPhos.TM..

Abstract: A process is disclosed for making a crystallized formulation for staining nuclear DNA to be used in flow cytometery. The crystallized DNA staining formulation and a kit containing the crystallized DNA staining formulation are provided. The staining formulation contains a mixture of a dye, a ribonuclease and a non-ionic hydrophilic surfactant. The process for preparing the formulation comprises mixing the dye, the enzyme and the surfactant and evaporating the mixture under vacuum to form a crystallized residue. The kit comprises the crystallized DNA staining formulation packed in single dose vials, each vial being sufficient for one analytic determination. Optionally, the kit may contain additional vials comprising a diluent used to reconstitute the crystallized residue at the time of use.

Abstract: An assay for activated factor VII (factor VIIa) has been developed using truncated tissue factor (tTF), a soluble mutant form of tissue factor (TF) that retains the cofactor function of TF toward factor VIIa. Unlike full-length TF, however, tTF appears not to support the conversion of factor VII to VIIa. As a result, the tTF assay for factor VIIa is free from interference from factor VII in the plasma and is therefore specific for factor VIIa. The assay is much simpler than existing assays, because it is a single-stage clotting assay performed almost identically to a prothrombin time (PT) assay. It is also considerably more sensitive than current assays for factor VIIa in plasma. Since the tTF assay is calibrated against a factor VIIa standard, it yields an absolute concentration of factor VIIa in ng/ml.

Abstract: An assay and test kit of the determination of LKM-1 autoantibodies in test samples suspected of containing anti-LKM-1 autoantibodies. The method uses a solid phase which preferably is a microparticle. The method is standardized and can be performed in automated systems, allowing quantitation of the amount of anti-LKM antibody in test samples.

Abstract: The present invention concerns a method to catalyze reporter deposition to improve detection or quantitation of an analyte in a sample by amplifying the detector signal which comprises reacting an analyte dependent enzyme activation system with a conjugate consisting of a detectably labeled substrate specific for the enzyme system, said conjugate reacts with the analyte dependent enzyme activation system to form an activated conjugate which deposits substantially wherever receptor for the activated conjugate is immobilized, said receptor not being reactive with the analyte dependent enzyme activation system.In another embodiment the invention concerns an assay for detecting or quantitating the presence or absence of an analyte in a sample using catalyzed reporter deposition to amplify the reporter signal.

Abstract: According to the process of the invention, sets of restriction fragments from the DNA of an individual are prepared, these restriction fragments being separated by size. Probes are prepared by enzymatic hydrolysis of DNA from a genome bank of the species to which this individual belongs, separation of the fragments obtained as a function of their size, labeling these probes and placing them in contact, under hybridization conditions, with the aforementioned sets of restriction fragments. Finally, selection is made of the probes (capable of hybridizing with said set of restriction fragments) which do not give hybridization profiles identical to those obtained with known probes recognizing variable number tandem repeat regions. Applications for the probes thus obtained include, particularly, processes for identifying an individual, consanguinity testing, and investigating the origin of a seed.

Abstract: Novel light producing 1,2-dioxetanes are described of the formula ##STR1## wherein ArOX is an aryl ring substituted with an X oxy group and A are passive organic groups which allow the 1,2-dioxetane to produce light when triggered by removing X. X is a chemically labile group which is removed by an activating agent. The 1,2-dioxetane compounds can be triggered to produce light at room temperatures.

Abstract: Provide herein is a substantially pure Lys-gingipain complex preparation, Lys-gingipain being characterized as having an apparent molecular mass of 105 kDa as estimated by sodium dodecyl sulfate polyacrylamide gel electrophoresis, where sample is prepared without boiling, said Lys-gingipain having amidolytic and proteolytic activity for cleavage after lysine residues and having no amidolytic and/or proteolytic activity for cleavage after arginine residues, wherein the amidolytic and/or proteolytic activity is inhibited by TLCK, cysteine protease group-specific inhibitors including iodoacetamide and iodoacetic acid, wherein the amidolytic and/or proteolytic activity of said Lys-gingipain is not sensitive to inhibition by leupeptin, antipain, trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane, serine protease group-specific inhibitors including diisopropylfluorophosphate and phenylmethyl sulfonylfluoride, and antibodies specific for the Lys-gingipain protein complex and its catalytic component, methods for p

Abstract: A method of identifying compounds or molecules which alter (enhance or inhibit) stimulation of kinase activity of pre-MPF and, thus, alter (enhance or inhibit) activation of MPF and entry into mitosis. The present method thus makes it possible to identify compounds or molecules which can be administered to regulate the cell cycle; such compounds are also the subject of this invention.

Abstract: A method for sequencing a strand of DNA, including the steps of: providing the strand of DNA; annealing the strand with a primer able to hybridize to the strand to give an annealed mixture; incubating the mixture with a deoxyribonucleoside triphosphate, a DNA polymerase, and a chain terminating agent under conditions in which the polymerase causes the primer to be elongated to form a series of DNA products differing in length of the elongated primer, each DNA product having a chain terminating agent at its elongated end; the number of each DNA product being approximately the same for substantially all DNA products differing in length from 1 to 20 bases.

Abstract: Fluorescently labelled dideoxynucleoside triphosphates are widely used as chain-terminators in Sanger dideoxy sequencing operations. But these unincorporated dye-terminators migrate in an electrophoresis gel and obscure the desired sequence ladder. This invention provides a method and a kit for modifying the unincorporated dye-terminators, e.g. by removal of a 5'-triphosphate group by chemical or enzymatic means e.g. by use of a phosphatase enzyme.

Abstract: The invention pertains to dipstick immunoassay devices. The device comprises a base member and a single, combined sample contact zone and test zone, wherein the test zone incorporates the use of symbols to detect analytes in a sample of biological fluid. A first immunological component, an anti-immunoglobulin capable of binding to an enzyme-labeled antibody, is immobilized in a control indicia portion. A second immunological component, capable of specifically binding to a target analyte which is bound to the enzyme-labeled antibody to form a sandwich complex, is immobilized in a test indicia portion. The enzyme-labeled antibody produces a visual color differential between a control indicia portion and a non-indicia portion in the test zone upon contact with a substrate.

Abstract: Enzyme activity is measured promptly with a high accuracy by introducing an enzyme, the activity of which is to be measured, into a column comprising a hollow tube packed with a filler comprising a support and a substrate that can be recognized by the enzyme, which is immobilized on the support, and measuring the amount of the obtained decomposition product of the substrate.

Abstract: The invention relates to a method for assaying homocysteine in a sample such as blood, plasma or urine, which comprises the steps of contacting the sample with a homocysteine converting enzyme and at least one substrate for the enzyme other than homocysteine, and without chromatographic separation, assessing a non-labelled analyte selected from a homocysteine co-substrate and the homocysteine conversion products of the enzymic conversion of homocysteine by said enzyme.

Abstract: A process using hydrothermally synthesized porous kaolinite as a carrier for use in a bioreactor. A carrier-biocatalyst composite body for use in a bioreactor, includes the synthesized kaolinite as a carrier and a biocatalyst fixed onto the synthesized kaoline. A bioreactor system includes a bioreactor vessel, and such a carrier-biocatalyst composite body placed in the bioreactor vessel.

Abstract: A method for measuring the amount of adenylate cyclase without the use of radioactive reagents is provided. The method comprises combining a sample of physiological material containing an amount of cAMP with (a) a mixture of enzymes effective to eliminate any other endogenous adenine nucleotides which may be present in the sample; and (b) an amount of alkaline phosphatase effective to eliminate any glucose-6-phosphate present in the sample. The cAMP present in said sample is then converted to AMP and the amount of AMP measured, which may then be correlated to the amount of cAMP and AC present in the sample.

Abstract: Monoclonal antibodies highly specific for human bone alkaline phosphatase, especially in the presence of human liver alkaline phosphatase, and their use in assays for human bone alkaline phosphatase are disclosed. A kit using the antibodies in an assay for human bone alkaline phosphatase is also disclosed.

Abstract: A method of enzymatic analysis utilizing a color-development signal amplification system associated with enzymatic cycling of NAD-NADH interconversion in the presence of dehydrogenase and its substrate, wherein the dehydrogenase is selected from the group consisting of alcohol dehydrogenase derived from Zymomonas and amino acid dehydrogenase derived from thermophilic microorganisms is disclosed. The use of alcohol dehydrogenase derived from Zymomonas provides an extremely higher detection sensitivity than that in the conventional method. The use of amino acid dehydrogenase derived from thermophilic microorganisms improves reliability of the method for a longer period of time than that in the conventional method.

Abstract: The invention is a non-invasive diagnostic test which is performed on the surface of the skin. This test indicates skin cholesterol levels which can provide information about the extent of aortic atherosclerosis. The invention also relates to reagents in the form of affino-enzymatic test compounds for use in the diagnostic test.

Abstract: The present invention concerns a method to catalyze reporter deposition to improve detection or quantitation of an analyte in a sample by amplifying the detector signal which comprises reacting an analyte dependent enzyme activation system with a conjugate consisting of a detectably labeled substrate specific for the enzyme system, said conjugate reacts with the analyte dependent enzyme activation system to form an activated conjugate which deposits substantially wherever receptor for the activated conjugate is immobilized, said receptor not being reactive with the analyte dependent enzyme activation system.In another embodiment the invention concerns an assay for detecting or quantitating the presence or absence of an analyte in a sample using catalyzed reporter deposition to amplify the reporter signal.

Abstract: The present invention is directed to the assay and purification of proteins, and particularly to the non-radioactive assay and purification of protein kinases, phosphatases and protease by incubating the enzyme with a substrate modified peptide to form a product modified peptide under conditions where the enzyme is active. The product modified peptide and substrate modified peptide are then separated, and the product modified peptide is measured. The present invention is also directed to kits and bioreagents for performing the assays.

Abstract: The CTD kinase of sporozoan parasites displays a specificity distinct from the analogous activity in mammalian cells. Methods of diagnosing blood borne Plasmodium parasites, and of testing the susceptibility of Plasmodium parasites to anti-malarial drugs, are based on this specificity.

Abstract: The method is directed to assaying for biological components in a sample comprising step (A) generating an oxidase substrate in the presence of an amphoteric surfactant and in the absence of ferrocyanide; (B) initially generating hydrogen peroxide through said oxidase reaction on said substrate of oxidase with subsequent detection of the generated hydrogen peroxide using peroxidase and a color developer capable of being oxidized in the presence of amphoteric surfactant and ferrocyanide; and (C) correlating the amount of color developed to the amount of biological components in the biological sample. Even when the biological component to be detected is present in a very small amount, the interference of bilirubin can be eliminated in assays of biological components in which peroxidase generated from an enzymatic reaction is detected using peroxidase and a color developer capable of being oxidized.

Abstract: Disclosed is a method of isolating a ligand from a sample, the method including: providing a hybrid molecule including the receptor for the ligand covalently bonded to the first member of a specific binding pair; contacting sample with the hybrid molecule to form an affinity complex between the ligand and the hybrid molecule; and isolating the affinity complex using the second member of the specific binding pair. Also disclosed is a c-kit ligand, nucleic acid encoding such a ligand, and recombinant cells containing such nucleic acid. The c-kit ligand may be used to stimulate hematopoietic cell growth.

Abstract: An assay method, compositions and test kits using a hydroxyaryl cyclic diacylhydrazide is described. A hydrogen peroxide and peroxidase enzyme. The preferred compositions incorporate enhancer compounds and a chelating agent which suppresses light production prior to addition of a peroxidase enzyme. The assay method can test for a peroxidase enzyme, a peroxide or can be used in immunoassays and probe assays.

Abstract: The invention relates to polymeric compositions useful in analyte determination. The compositions contain a polymer, a reagent system for analyte determination, and an extender. The last component alleviates tackiness in the composition, and thus reduces damage in preparation of test apparatus. Mica is the particularly preferred extender.

Abstract: Enzymatically clearable chemiluminescent 1,2-dioxetane compounds capable of producing light energy when decomposed, substantially stable at room temperature before a bond by which an enzymatically clearable labile substituent thereof is intentionally cleaved, are disclosed. These compounds can be represented by the formula: ##STR1## wherein: X and X.sup.1 each represent, individually, hydrogen, a hydroxyl group, a halo substituent, an unsubstituted lower alkyl group, a hydroxy (lower) alkyl group, a halo (lower) alkyl group, a phenyl group, a halophenyl group, an alkoxyphenyl group, a hydroxyalkoxy group, a cyano group or an amide group, with at least one of X and X.sup.1 being other than hydrogen; and R.sub.1 and R.sub.2, individually or together, represent an organic substituent that does not interfere with the production of light when the dioxetane compound is enzymatically cleaved and that satisfies the valence of the dioxetane compound's 4-carbon atom, with the provisos that if R.sub.1 and R.sub.

Abstract: A rapid radioactive method of measuring enzymatic activity of a protein kinase is disclosed. The method is an improvement to existing methodology which involves phosphorylating a peptide substrate using .sup.32 P-ATP, adsorbing the phosphorylated peptide to a solid phase, washing the phase to remove non-adsorbed .sup.32 P-ATP, and measuring the radioactivity of the phosphorylated peptide adsorbed to the phase. The disclosed improvement uses a membrane as the solid phase and positions the membrane within a chamber to separate the chamber into a first and second region. Washing is accomplished with centrifugal force; the washed solution being forced through the membrane from the first region into the second region.

Abstract: A kit for carrying out an immunoenzymological assay for protein, containing a diagnostic reagent consisting of a hybrid protein comprising, in order, a polypeptide P1 which is an N-terminal portion of mature alkaline phosphatase comprising from 6 to 28 of the N-terminal amino acids of said mature alkaline phosphatase, joined to a polypeptide P2, capable of interacting with an antibody or antigen joined to a polypeptide P3 which is selected from the group consisting of a) the C-terminal remaining amino acids of said mature alkaline phosphatase, b) the mature sequence of alkaline phosphatase, and c) enzymatically active fragments of b).

Abstract: A method for assaying nucleic acids or similar compounds comprises binding a sample such as a nucleic acid to phosphatase; reacting the phosphatase with a 3-hydroxy-2-naphthoic acid-2'-phenyl anilide phosphate; irradiating the reaction product with an excited light; and detecting fluorescence emitted therefrom.

Abstract: A system or kit for analysis of phosphatase comprising a reagent I containing NADP or NADPH as a main component, and a reagent II containing components for color-development signal amplification utilizing NAD-NADH interconversion, wherein at least one of the hydrogen donors participating in the NAD-NADH interconversion is included in reagent I, but not in reagent II, which eliminates the necessity that the reagent II is further separated into two parts. This enables the preparation of reagent II as a mixture which can be readily used and which can be preserved for a long period of time without "reagent blank" occurring. Additionally, the present kit eliminates the necessity of mixing reagents I and II immediately before actual use and thus permits periodical analysis of samples with a reagent having the same components for long time periods with reliable and uniform sensitivity.

Abstract: A colorimetric enzymic analytical test element comprises a support and a plurality of reaction zones incorporating a dried enzyme composition, dyestuff and reagent adapted to provide a substantial dosage independent colorimetric display when the concentration of an analyte applied to the elements exceeds a predetermined value.

Abstract: A method of measuring enzymatic activity of a protein kinase is disclosed. The method is an improvement to existing methodology which involves phosphorylating a peptide substrate, adsorbing the phosphorylated peptide to a solid phase, washing the phase to remove non-adsorbed constituents, and measuring the amount of phosphorylated peptide adsorbed to the phase. The disclosed improvement uses a membrane as the solid phase and positions the membrane within a chamber to separate the chamber into a first and a second region. Washing is accomplished with centrifugal force; the washed solution being forced through the membrane from the first region into the second region. Radioactive and non-radioactive assays are disclosed. The latter uses a support containing the Fe.sup.+3 ion chelated to the support through imminodiacetic acid groups. The peptide contains a dye and measurement is spectrophotometric or fluorometric.

Abstract: Monoclonal antibodies highly specific for human bone alkaline phosphatase, especially in the presence of human liver alkaline phosphatase, and their use in assays for human bone alkaline phosphatase are disclosed. A kit using the antibodies in an assay for human bone alkaline phosphatase is also disclosed.

Abstract: Aryl N-alkylacridanthiocarboxylate compounds which produce chemiluminescence. The compounds produce light with peroxide and peroxidase. The compounds are used as a substrate in assays for various analytes.

Abstract: The present invention discloses novel compounds useful as alkaline phosphatase inhibitors and therapeutic agents. Preferably, the novel compounds are useful as selective inhibitors of human alkaline phosphatases as opposed to Escherichia coli alkaline phosphatases. The novel compounds can also be used as cancer therapeutic agents, anti-depressive agents, anti-anergic agents, and antihelminthic agents. The novel compounds have the following general formula: ##STR1## wherein R' is an aryl, aryl ether, aryl thioether, aromatic heterocyclic, aromatic heterocyclic thioether, or aromatic heterocyclic ether group. More preferably, R' is a phenyl or a pyridine. Most preferably, R' is of the following formula: ##STR2## 2-thiopyridine, or ##STR3## 2-oxypyridine. R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.1 ', R.sub.2 ', R.sub.3 ', R.sub.4 ', and R.sub.5 ' can be the same or different, and at least one of which is selected from the group consisting of: H, C.sub.1 -C.sub.6 alkyl, halo C.sub.1 -C.sub.6 alkyl, phenyl, C.

Abstract: A dry immunoassay analytical element, for assaying a ligand, comprising a support bearing:(a) a labeled ligand zone;(b) a spreading zone; and(c) a receptor zone containing a fixed concentration of an immobilized receptor for the ligand and the labeled ligand and the receptor is covalently bonded to polymeric beads having a diameter in the range of 0.1 to 5 .mu.m;characterized in that the element contains a compound containing a vanadium IV (V.sup.+4) ion and the zones can be in the same or separate layers.

Abstract: A method for the identification of related polypeptides, using an function-based selection criterion, is disclosed. A novel human phosphatase, and nucleotide sequences coding therefor, identified by the aforementioned method and designated VHR, is also disclosed.

Abstract: A method and multilayer analytical element for the determination of catechol and catechol generating substances such as salicylate is described. A series of enzymatic conversions involving tyrosinase is used to convert catechol to o-quinone and the latter to convert a leuco dye to a colored dye.