Abstract: Methods for enhancing luminescence of a luciferase (BFP-aq) with fluorescence activity derived from a calcium-binding photoprotein are provided. To a luciferase solution with fluorescence activity that contains an apoprotein, a calcium-binding photoprotein, which is constituted such that a coelenteramide or an analog thereof is coordinated inside, a coelenterazine that is the luminescent substrate of the luciferase or an analog thereof and a compound (e.g., imidazole etc.) having the function of removing an —NH-proton of the pyrazine ring of the imidazopyrazine skeleton in the coelenterazine or the analog thereof are added.
Abstract: The present invention provides a fast and efficient means for identifying kinetically stable proteins. As used herein the term “kinetically stable protein” means a protein that is trapped in a specific conformation due to an unusually high unfolding barrier that results in very slow unfolding rates. The present inventors are the first to discover the existence of a correlation between kinetic stability and SDS-induced denaturation. Thus, the invention provides methods for identifying kinetically stable proteins comprising the step of testing the proteins for resistance to denaturation by SDS. In one embodiment, SDS-polyacrylamide gel electrophoresis (SDS-PAGE) is one simple method for quickly identifying and selecting kinetically stable proteins.
Abstract: The present invention includes a device and associated analytical method to use for the sensitive detection and accurate, rapid determination of acrylamide in food substances. The present invention also relates to the use of a kit device and associated analytical method in which a user can quickly and easily ascertain the amount of acrylamide in food substances with ease and in any location, including a non-laboratory environment. Such detection device and method may be comprised of a sample collection area on which a sample of food, after being mixed in a solution, is placed for example on the substrate of a biochip that includes an enzyme that along with a co-enzyme or form of energy or catalyst, facilitates the conversion of either acrylamide to acrylonitrile or the conversion of acrylamide to ammonia.
Abstract: The present invention relates generally to enzyme assays and more particularly to a rapid, sensitive, reliable and robust homogeneous assay for acetyl CoA carboxylase activity comprising a coupled enzyme assay in a scintillation proximity format suitable for high-throughput screening. In one aspect, the assay couples ACC and FAS and the product, palmitic acid is then detected by scintillation counting. The invention also relates to the identification of modulators of ACC activity.
Abstract: A reagent for measuring an alanine aminotransferase activity comprising L-alanine, 2-oxoglutaric acid, lactate dehydrogenase, and reduced nicotinamide adenine dinucleotide, characterized by further comprising a substance having an activity of inhibiting lactate dehydrogenase activity is disclosed. Further, a method for measuring an alanine aminotransferase activity, characterized by bringing a sample to be analyzed, which may contain alanine aminotransferase, into contact with L-alanine, 2-oxoglutaric acid, lactate dehydrogenase, reduced nicotinamide adenine dinucleotide, and a substance having an activity of inhibiting a lactate dehydrogenase activity is disclosed. According to the reagent and method, an increase in the reagent blank reaction, i.e., an increase in the initial absorbance, can be suppressed.
Abstract: A modified limulus test procedure is provided to measure the lipopolysaccharide (LPS), lipid A, and/or glucan reactivity in a biological liquid. The test procedure is independent of the anti-limulus factors present in the sample. The procedure involves strong oxidative treatment of the biological liquid sample.
Abstract: An improved method for detecting a target biomolecule directly in a polyacrylamide gel in which it has been separated from other substances. The improvement resides in, prior to binding the target to a probe and while the target biomolecule remains in the gel, (1) immersing the gel in a water miscible, aqueous extracting medium to shrink the gel by at least about ten percent and then (2) washing the gel with water to restore the gel to substantially its original size.
Abstract: A method of identifying compounds capable of modulating trans-glutaminase (TG) kinase activity is described. The method involves adding a test compound to a mixture of TG and a suitable TG kinase substrate, incubating the mixture under conditions promoting TG kinase activity, and determining if the test compound activates or modulates TG kinase activity as indicated by greater than expected TG-mediated phosphorylation of the TG kinase substrate or if the test compound inhibits TG kinase activity as indicated by lower than expected TG-mediated phosphorylation of the TG kinase substrate as compared to a control comprising TG and a suitable TG kinase substrate incubated under conditions promoting TG kinase activity.