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Additional info for Bioluminescence and Chemiluminescence
The product is cooled, and small aliquots are poured onto crushed ice (no excess liquid present) and mixed to ensure good yields. The resultant precipitate is filtered and extracted with benzene. This extract is filtered, and petroleum ether is added slowly to give a colorless precipitate. This yields approximately 30 g of 2-cyano-6-methoxybenzothiazole (V). p. 128°-130°). Preparation of 2-Cyano-6-hydroxybenzothiazole (VI) Removal of the methyl group at position 6 is accomplished in a pyridine hydrochloride melt at 200°-210 °.
8 show that luciferin is not adsorbed to BioGel P-10. Hence it is not separated from ATP on this material. The adsorption to Sephadex G- 10 is greatly increased at pH 3. 9 This has been used for concentrating and purifying luciferin from analytical reaction mixtures containing extract of 1 g of firefly tails per liter. Only little of the luciferin is used up in the analytical reaction, and the reaction mixtures may be stored frozen for years. 5 × 20 cm for 1 liter), and elution of luciferin with neutral buffer.
Ifa small amount of pure luciferin is available, fluorescence excitation and emission spectra of pure versus synthetic luciferin can be compared and provides another convenient means of estimating purity. If pure luciferin is not available, fluorescence excitation and emission spectra can be corrected for lamp output and phototube sensitivity and compared to pub10M. DeLuca, L. Brand, T. A. Cebula,H. H. Seliger,and A. F. Makula,J. Biol. Chem. 246, 6702 (1971). 11H. H. Seligerand W. D. McElroy,Arch.