Chemiluminescent probe

Lucas, M., and Solano, F. (1992). Coelenterazine is a superoxide anion-sensitive chemiluminescent probe its usefulness in the assay of respiratory burst in neutrophils. Anal. Biochem. 206 273-277. 

Nakano, M., Sugioka, K., Ushijima, Y., and Goto, T. (1986). Chemiluminescence probe with Cypridina luciferin analogue, 2-methyl-6-phenyl-3,7-dihydroimidazo[l,2-a]pyrazine-3-one, for estimating the ability of human granulocytes to generate superoxide anion. Anal. Biochem. 159 363-369. 

Nishinaka, Y., et al. (1993). A new sensitive chemiluminescence probe, L-012, for measuring the production of superoxide anion by cells. Biocbem. Biophys. Res. Commun. 193 554—559. 

Teranishi, K. (2003). Cyclodextrin-bound 6-(4-methoxyphenyl)imidazo[l,2-a]-pyrazin-3(7H)-one as chemiluminescent probe for superoxide anions. ITE Letters on Batteries, New Technologies and Medicine 4 201-205. 

Teranishi, K., and Shimomura, O. (1997b). Coelenterazine analogs as chemiluminescent probe for superoxide anion. Anal. Biochem. 249 37-43. 

Witko-Sarsat, V., Nguyen Anh Thu, Knight, J., and Descamps-Latscha, B. (1992). Pholasin a new chemiluminescent probe for the detection of chloramines derived from human phagocytes. Free Radic. Biol. Med. 13 83-88. 

Keshavarzian, A., Sedghi, S., Kanofiky, J.R, List, T., Robinson, C., Ibrahim, C. and Winship, D. (1992c). Excessive production of reactive oxygen metabolites by inflamed colon analysis by chemiluminescence probe. Gastroenterology 103, 177-185. 

Suematsu, M., Suzuki, M., Kitahora, T. et al. (1987a). Increased respiratory burst of leukocytes in inflammatory bowel disease - the analysis of free radical generation by using chemiluminescence probe. J. Clin. Lab. Immunol. 24, 125-128. 

Li X, Zhang G, Ma H et al (2004) 4, 5-Dimethylthio-4 -[2-(9-anthryloxy)ethylthio]tetra-thiafulvalene, a highly selective and sensitive chemiluminescence probe for singlet oxygen. J Am Chem Soc 126 11543-11548... 

The second label also may be a fluorescent compound, but doesn t necessarily have to be. As long as the second label can absorb the emission of the first label and modulate its signal, binding events can be observed. Thus, the two labeled DNA probes interact with each other to produce fluorescence modulation only after both have bound target DNA and are in enough proximity to initiate energy transfer. Common labels utilized in such assay techniques include the chemiluminescent probe, N-(4-aminobutyl)-N-ethylisoluminol, and reactive fluorescent derivatives of fluorescein, rhodamine, and the cyanine dyes (Chapter 9). For a review of these techniques, see Morrison (1992). 

Scheme 7 shows that the method of sequential arylation with a high selectivity, using 2-phenylimidazole motif, proves to be applicable for pharmaceuticals and fluorescent and chemiluminescent probes.85 The direct 4-arylation of free 2-phenylimidazole is achieved with iodoarenes as the aryl donors in the presence of palladium catalyst (Pd/PPh3) and MgO as the base. A complete switch from G4 to C2 arylation is accomplished using a ruthenium catalyst [CpRu(PPh3)2Cl] and Gs2G03. 

Acridinium ester—labeled chemiluminescent probes have been utilized to detect the specific protein-coding transcripts and to distinguish between transcripts that code for the 190-kDa protein and the two closely related 210-kDa proteins. The assay is called the hybridization protection assay (D3). In this assay, RNA isolated from the patient s white blood cells is first amplified by PCR. The amplified product is incubated with the chemiluminescent probe. The unhybridized probe is removed by selective hydrolysis in sodium tetraborate buffer, containing surfactant Triton X-100 at pH 8.5, in an incubation step at 60°C for 6 min. After the sample is cooled to room temperature, the chemiluminescence of the hybridized probe is measured in a luminometer. The procedure is reported to detect one leukemic cell in a population of a million or more normal cells. It is also rapid, requiring less than 30 min. Its reliability has been attested to by correlation with results obtained on karyotypic and Southern blot analysis (D3). 

Gl. Gold, B., Radu, D., et al., Diagnosis of fragile X syndrome by Southern blot hybridization using a chemiluminescent probe A laboratory protocol. Mol. Diagn. 5(3), 169-178 (2000). 

Dioxetanes, labeled with triggers sensitive to the alkaline-phosphatase enzyme, serve as highly sensitive chemiluminescent probes in numerous bioassays. Current applications include immunoassays, membrane-based detection of proteins and nucleic acids, and microplate-based and array-based nucleic-acid detection. ... 

The synthesis of A-methylacridan dioxetane derivatives would be very promising for analytical application as chemiluminescence probes. Despite the high quantum yields... 

Several other examples of 1,2-dioxetane derivatives containing easily oxidizable groups have been reported and the high singlet quantum yield observed in their decomposition was attributed to the occurrence of the intramolecular CIEEL sequence Based on this concept, Schaap and coworkers have introduced the concept of induced chemiluminescence, which is very relevant for investigations into firefly luciferin bioluminescence and has led to the development of chemiluminescent probes widely used in immunoassays (Section N. 

Chemiluminescent probes enable highly sensitive quantitative analysis of proteins blotted from electrophoretic gels onto a supporting matrix. For a quantitative comparison, it is important to be able to correct for introduced variables such as antibody titre, temperature, substrate etc. Comparison of blots completed on different days requires a chemiluminescent standard. The situation is more complex with 2D gels, where only one sample per gel/blot is used. A method has been published for preparing a chemiluminescent standard for quantitative comparison of 2D Western blot (89). 

Chemiluminescent probes based on the dioxetane moiety are now being developed for the detection of cholinesterase activity <2002JA4874>. In this particular case, the nucleophilic thiol generated from hydrolysis of acetylthiocholine iodide triggers the formation of the thiolate 61 and by-product 62 through nucleophilic attack on the disulfide bond of 60 (Scheme 14). 

Methodologically, both techniques are based on the digestion of the genomic DNA with restriction endonucleases, separation of the fragments by electrophoresis, followed by X transfer to a nylon membrane and finally, detection by hybridization with either a radioactive or chemiluminescent probe. This technique is known as Southern blot analysis. 

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