Abstract: Provided herein are methods for detecting and quantifying succinate in a sample. Also provided are methods for detecting and quantifying 2-oxoglutarate oxygenase enzyme and/or 2-oxoglutarate oxygenase activity in a sample and methods for screening for modulators of 2-oxoglutarate oxygenase activity.

Abstract: Provided herein are compositions and methods for the assembly of a bioluminescent complex from two or more non-luminescent (e.g., substantially non-luminescent) peptide and/or polypeptide units. In particular, bioluminescent activity is conferred upon a non-luminescent polypeptide via structural complementation with another, complementary non-luminescent peptide.

Abstract: Provided herein are methods for detecting and quantifying succinate in a sample. Also provided are methods for detecting and quantifying 2-oxoglutarate oxygenase enzyme and/or 2-oxoglutarate oxygenase activity in a sample and methods for screening for modulators of 2-oxoglutarate oxygenase activity.

Abstract: Provided herein are compositions and methods for the assembly of a bioluminescent complex from two or more non-luminescent (e.g., substantially non-luminescent) peptide and/or polypeptide units. In particular, bioluminescent activity is conferred upon a non-luminescent polypeptide via structural complementation with another, complementary non-luminescent peptide.

Abstract: Provided herein are compositions and methods for the assembly of a bioluminescent complex from two or more non-luminescent (e.g., substantially non-luminescent) peptide and/or polypeptide units. In particular, bioluminescent activity is conferred upon a non-luminescent polypeptide via structural complementation with another, complementary non-luminescent peptide.

Abstract: Provided herein are methods for detecting and quantifying 5-hydroxymethylated cytosine bases in a DNA molecule.

Abstract: Luminescent detection of inorganic phosphate is carried out in an assay through the intermediary enzymatic production of ADP. ADP is converted to ATP which is used in a luminescent reaction. The assay can be used to monitor coupled enzyme reactions which use or generate inorganic phosphate and the modulation of such reactions.

Abstract: The invention provides compositions and methods to determine or detect the activity of enzymes, including phosphotransferases such as kinases (e.g., protein, lipid, and sugar kinases) and ATP hydrolases such as ATPases, e.g., HSP90, that employ ATP as a substrate and form ADP as a product by monitoring changes in ADP. Methods, kits and compositions for monitoring changes in metabolites such as ADP are disclosed.

Abstract: The invention provides provides compositions and methods to determine or detect the activity of enzymes, including phosphotransferases such as kinases (e.g., protein, lipid, and sugar kinases) and ATP hydrolases such as ATPases, e.g., HSP90, that employ ATP as a substrate and form ADP as a product by monitoring changes in ADP.

Abstract: The disclosure provides compositions and methods to determine or detect ADP or the activity of enzymes, including phosphotransferases such as kinases (e.g., protein, lipid, and sugar kinases) and ATP hydrolases such as ATPases, e.g., HSP90, that employ ATP as a substrate and form ADP as a product by monitoring changes in ADP.

Abstract: The present invention relates in general to cellular analysis tools and more particularly to methods for detecting or determining cyclic nucleotide concentrations in samples. Samples containing cyclic nucleotides may be contacted with a cyclic nucleotide-dependent protein kinase and a detection system which includes a substrate for the cyclic nucleotide-dependent protein kinase. The activities in cyclic nucleotide related pathways may be measured using the detection system.

Abstract: Methods and kits for detecting transferase activity in a sample by measuring ATP using a composition comprising an ATP-dependent bioluminescence-generating enzyme, a luminogenic molecule and one or more transferase quenching agents.

Abstract: The invention provides compositions and methods to determine or detect the activity of enzymes, including phosphotransferases such as kinases (e.g., protein, lipid, and sugar kinases) and ATP hydrolases such as ATPases, e.g., HSP90, that employ ATP as a substrate and form ADP as a product by monitoring changes in ADP.

Abstract: Methods and kits for detecting transferase activity in a sample by measuring ATP using a composition comprising an ATP-dependent bioluminescence-generating enzyme, a luminogenic molecule and one or more transferase quenching agents.

Abstract: Kits and peptide substrates for detecting transferase activity of a sample are provided. The kits include a substrate including a reporter compound, at least one of a phosphate group donor and a phosphate group acceptor, a buffer that supports enzymatic activity of the transferase, and a peptidase that cleaves a non-phosphorylated peptide substrate at a first rate and a phosphorylated peptide substrate at a second rate. The peptide substrates include a reporter compound and a first transferase substrate linked to the reporter compound on a first side of the reporter compound.

Abstract: A method for detecting alteration in a kinase or phosphatase reaction is provided. In the method, a test substance is contacted to a peptide substrate including a reporter compound and amino acids under conditions in which the kinase or phosphatase is active. The substrate is cleaved with a carboxypeptidase. Phosphorylation of the substrate affects cleavage by the carboxypeptidase. Output of the reporter compound is detected. The reporter compound exhibits a different output property when bound to at least one amino acid of the substrate when compared to when it is not bound to amino acids. Change in output compared to output of a control sample that has not been contacted with the test substance is a measure of the alteration in the kinase or phosphatase reaction.

Abstract: A method for detecting transferase activity of a sample includes contacting the sample with a substrate and at least one of a phosphate group donor and a phosphate group acceptor. The substrate includes a reporter compound and amino acids. A peptidase is added that cleaves a non-phosphorylated substrate at a first rate and a phosphorylated substrate and a second rate. The output of the reporter compound is detected. In a preferred embodiment, the transferase activity detected is a kinase activity. In another preferred embodiment, the transferase activity detected is a phosphatase activity. Also provided is a method of screening for alterations in a transferase reaction. Kits and peptide substrate are also provided for carrying out at least one of the methods of the invention.

Abstract: Methods for detecting transferase activity of a sample are provided. Transferase activity detected includes kinase and phosphatase activity. The method includes providing a substrate having a reporter compound conjugated thereto. A quantity of the substrate is added to a solution containing the sample. The sample is incubated with the substrate under conditions where the sample is active for a time sufficient for kinase or phosphatase activity to take place. An exopeptidase is added to the solution containing the sample. Output of the reporter compound is detected.

Abstract: The present invention relates in general to cellular analysis tools and more particularly to methods for detecting or determining cyclic nucleotide concentrations in samples.

Abstract: A method for detecting transferase activity of a sample includes contacting the sample with a substrate and at least one of a phosphate group donor and a phosphate group acceptor. The substrate includes a reporter compound and amino acids. A peptidase is added that cleaves a non-phosphorylated substrate at a first rate and a phosphorylated substrate and a second rate. The output of the reporter compound is detected. In a preferred embodiment, the transferase activity detected is a kinase activity. In another preferred embodiment, the transferase activity detected is a phosphatase activity. Also provided is a method of screening for alterations in a transferase reaction. Kits and peptide substrate are also provided for carrying out at least one of the methods of the invention.