By Michael J. Corey
This ebook highlights the functions of coupled bioluminescence assay recommendations to real-world difficulties in drug discovery, environmental and chemical research, and biodefense. It separates theoretical features 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 investigate toxicity, drug receptors, and enzymes. It covers functions in environmental research and biodefense, together with cytotoxicity, fertilizer and explosives research, and nerve agent and pesticide detection. this is often the preferable reference on coupled bioluminescent assays for chemists, biochemists, and molecular biologists.Content:
Chapter 1 advent (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 examine (pages 85–102):
Chapter five Coupled Bioluminescent Phosphatase Assays (pages 103–130):
Chapter 6 Acetylcholinesterase (pages 131–138):
Chapter 7 dimension 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 matters (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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Extra info for Coupled Bioluminescent Assays: Methods, Evaluations, and Applications
Subsequent work on less stable 1,2-dioxetanes led to the issuance of a broad patent in 1994 covering “Enzymatically cleavable chemiluminescent 1,2-dioxetane compounds capable of producing light energy when decomposed” (18). One of the major advances of this work was the provision of water-soluble 1,2-dioextane substrates. Other advantages of the new admantylidene dioxetane compounds, including a more rapid approach to steady-state light emission, were claimed. Alkaline phosphatase, ␤-galactosidase, and (by implication) esterases, acyl transferases, glycosidases, amidases, and a wide variety of other hydrolases were among the enzyme groups named as actual or potential assay targets for these chemiluminescent substrates.
NADH is much more stable to spontaneous oxidation than FMNH2 ; hence, the reaction series has evidently evolved to produce FMNH2 in situ when needed. 1 Mechanism of Action of Bacterial Luciferases Elucidation of the mechanism of bacterial luciferases has been a challenging problem, and many scientists have made contributions to our understanding of the very complex reaction series (49–53). Bacterial luciferases may be viewed as mixed-function oxidases that utilized molecular oxygen to oxidize two substrates in a single turnover: FMNH2 and the long-chain aldehyde, which is oxidized to a carboxylic acid.
Another advance was realized with the principle of electrochemiluminescence (ECL), which was also called “electrogenerated chemiluminescence” in early publications (19–21). This allows an even higher level of control of the timing and position of the chemiluminescent event. For example, the acridan ester 2 ,6 -difluorophenyl 10-methylacridan-9-carboxylate decays to an acridinium ester in response to an electrical impulse when an appropriate oxidant is present, and the decay event can be timed to occur following an interval subsequent to a diffusion event, enzyme activity, or addition of another reaction component (such as coated magnetic beads) (22).