Promega Corporation

James J Cali

Keith V Wood

Erika Hawkins

Dieter Klaubert

Wenhui Zhou

William Daily

Samuel Kin Sang Ho

John William Shultz

Poncho Meisenheimer

Martha A O'Brien

Michael P Valley

Sarah Duellman

Robert F Bulleit

Jolanta Vidugiriene

Richard Somberg

Susan Frackman

James Unch

Jianquan Liu

Michael Scurria

Brock Binkowski

Christopher Todd Eggers

Dongping Ma

Hui Wang

Donna M Leippe

Natasha Karassina

Mary Sobol

Peter Hofsteen

Jessica Kelts

Jessica Anderson

John W Schultz

Helene A Benink

Dana Mustafa

Samuel K Ho

Erica Hawkins

Martha O'BRIEN

Donna Leippe

Provided herein are materials and methods for performing bioluminescent assays using a bifunctional probe. In particular, the present disclosure provides compositions and methods for detecting and/or quantifying a biomolecule and/or assaying a cellular process associated with the biomolecule using a bifunctional probe capable of binding the biomolecule and generating a bioluminescent and/or fluorescent signal.

Compositions and methods for bioluminescent detection using multifunctional probes

Conjugates comprise a drug or probe, a cell binding agent, and a quinone-containing linker. The quinone-containing linker may be reduced intracellularly to trigger release of the drug or probe. These conjugates may selectively deliver a drug or probe to a site of action of interest for local release of the active drug or probe.

Quinone-containing conjugates

Methods, kits and compositions containing a mixture of D-luciferin and L-luciferin for light generation with luciferase are disclosed that have improved stability when stored over time. The mixture of D-luciferin and L-luciferin can be used to detect the presence or amount of ATP or of luciferase in a sample.

Stabilized formulation for luminescent detection of luciferase and nucleoside phosphates

Provided herein are methods for the real-time monitoring of an intracellular event or response. In particular, the methods provided herein monitor the conversion of a pro-substrate to a substrate for a protein sensor as a result of an intracellular event or response.

Real-time monitoring

A method to detect the presence or amount of at least one molecule in a sample which employs a derivative of luciferin or a derivative of a fluorophore is provided. Compounds and compositions for carrying out the methods of the invention are also provided.

Luminogenic and fluorogenic compounds and methods to detect molecules or conditions

The invention provides compounds, compositions, methods, substrates, and kits useful for analyzing the metabolic activity in cells, tissue, and animals and for screening test compounds for their effect on cytochrome P450 activity. In particular, a one-step and two-step methods using luminogenic molecules, e.g. luciferins or coelenterazines, that are cytochrome P450 substrates and that are also bioluminescent enzyme, e.g., luciferase, pro-substrates are provided. The present method further provides a method for stabilizing and prolonging the luminescent signal in a luciferase-based assay using luciferase stabilizing agents such as reversible lucifetrase inhibitors.

Luminescence-based methods and probes for measuring cytochrome P450 activity

A method to detect the presence of amount of at least one molecule in a sample which employs a derivative of luciferin or a derivative of a fluorophore is provided. Compounds and compositions for carrying out the methods of the invention are also provided.

The present invention provides compounds and methods for assaying redox state of metabolically active cells and methods for assaying enzyme activity and/or metabolite level by coupling to redox defining co-factor NAD(P)/NAD(P)H measurement.

Compounds and methods for assaying redox state of metabolically active cells and methods for measuring NAD(P)/NAD(P)H

Provided are labeling reagents and methods of using the reagents for cell uptake measurements. The labeling reagents can be quinone-masked probes including fluorophores and/or luminophores.

Quinone-masked probes as labeling reagents for cell uptake measurements

A method and kit are provided for enhancing the tolerance of an assay reagent to compounds in an assay sample, the assay reagent including a luciferase enzyme. The method includes contacting the luciferase with a tolerance enhancement agent in an amount sufficient to substantially protect luciferase enzyme activity from interference of the compound and minimize interference by at least about 10% relative to an assay not having tolerance enhancement agent.

Luciferase-based assays

The invention provides compounds, compositions, methods, substrates, and kits useful for analyzing the metabolic activity in cells, tissue, and animals and for screening test compounds for their effect on cytochrome P450 activity. In particular, a one-step and two-step methods using luminogenic molecules, e.g. luciferins or coelenterazines, that are cytochrome P450 substrates and that are also bioluminescent enzyme, e.g., luciferase, pro-substrates are provided. The present method further provides a method for stabilizing and prolonging the luminescent signal in a luciferase-based assay using luciferase stabilizing agents such as reversible luciferase inhibitors.

Growing community of inventors

Verona, WI, United States of America

James J Cali