By Michael J. Corey
This booklet highlights the functions of coupled bioluminescence assay thoughts to real-world difficulties in drug discovery, environmental and chemical research, and biodefense. It separates theoretical elements from the utilized sections in a transparent and readable method. Coupled Bioluminescent Assays, explains the makes use of of CB applied sciences throughout drug discovery to research toxicity, drug receptors, and enzymes. It covers purposes in environmental research and biodefense, together with cytotoxicity, fertilizer and explosives research, and nerve agent and pesticide detection. this can be the most effective reference on coupled bioluminescent assays for chemists, biochemists, and molecular biologists.Content:
Chapter 1 creation (pages 1–23):
Chapter 2 Coupled Bioluminescent Reactions in perform (pages 24–55):
Chapter three Coupled Bioluminescent Cytotoxicity Assays (pages 57–84):
Chapter four The function of Coupled Bioluminescent Assays in Kinase Screening and learn (pages 85–102):
Chapter five Coupled Bioluminescent Phosphatase Assays (pages 103–130):
Chapter 6 Acetylcholinesterase (pages 131–138):
Chapter 7 size of Nitric Oxide Synthase job through Coupled Bioluminescence (pages 139–149):
Chapter eight The Coupled Bioluminescent Pyrophosphorolysis Assay (pages 150–159):
Chapter nine Coupled Luminescent Assays of G?Protein?Coupled Receptors (pages 160–190):
Chapter 10 Coupled Bioluminescent Protease Assays (pages 191–200):
Chapter eleven Coupled Luminescent Assays related to Aequorin (pages 201–208):
Chapter 12 Coupled Bioluminescent Reporter Assays (pages 209–217):
Chapter thirteen Coupled Bioluminescent Assays: Regulatory issues (pages 218–240):
Chapter 14 Coupled Bioluminescent decision of Bioburden and Sterility (pages 241–246):
Chapter 15 Environmental functions of Coupled Bioluminescent Assays (pages 247–258):
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Additional resources for Coupled Bioluminescent Assays: Methods, Evaluations, and Applications
A notable example is pyrophosphate, which can not only serve as a competitive inhibitor of ATP but also help to initiate the backward reaction of luciferase (generation of ATP in the presence of light, pyrophosphate, and AMP), greatly complicating interpretation of the assay results. The actual species that participates in the luminescent reaction is not ATP, but a complex of ATP with the Mg2+ ion. The cation is essential, and reagents that chelate or sequester this ion, such as EDTA and its various cousins or apoenzymes, may shut down the light reaction.
The operator should perform an instrument check routine every day when critical work is planned, and it should be possible to initiate this routine with minimal effort and complexity. The software should enable selection of any rectangular subset of the plate wells for reading. Along with this feature, a very important capability of both software and instrument is the ability to predict, or at least test, the exact cycle time of both the individual reads and the entire experimental run. This stands in marked contrast, for example, to the case of ELISA readouts, in which time-critical changes are usually not occurring within seconds.
Creatine phosphate is the proximal backup molecule that can provide ATP units with great rapidity through the action of creatine kinase on creatine phosphate. 11 Lundin’s coupled bioluminescent assay for creatine kinase activity. This, the first coupled bioluminescent assay published, is based on the simple concept that ATP evolved by the kinase-catalyzed reaction can be used by luciferase to produce visible light in proportion to the concentration of the kinase. Creatine kinase catalyzes the transfer of the phosphate group from creatine phosphate to ADP, generating ATP and enabling the light reaction.
Coupled Bioluminescent Assays: Methods, Evaluations, and Applications by Michael J. Corey