Chemiluminescence probe MCLA

Chemiluminescence was measured in triplicate at 23 "C under moderate agitation by use of Luminescence Reader BLR-201 (Aloka Co. Ltd Tokyo Japan). In a glass test tube, 100 /xL of chemiluminescent probe suspension, 450 fiL of distilled and deionized water, 100 /iL of 10-fold concentrated PBS and 250 fiL of 1 U/mL solution of xanthine oxidase from butter milk (XOD) were mixed about 30 s after the start of CL measurement following the set of the test tube in the instrument, 100 fiL of 1.67 mM hypoxanthine (HPX) solution was injected into the test tube through a rubber cap by a syringe. The initial concentration of MCLA or FCLA was 1 fiM. The pH of the reaction mixture was 7.3 0.1. 

In this study I have developed a near-infrared chemiluminescenct probe 9 for the detection of superoxide anions. First, the present study proved that CRET with MCLA and indocyanine molecules is useful to generate near-infrared light. Second, the near-infrared chemiluminescent probe 9 is easily synthesized. Finally, the suitability of 9 for the detection of superoxide anions was demonstrated. 

Wang J, Xing D. Detection of vitamin C-induced singlet oxygen formation in oxidized LDL using MCLA as a chemiluminescence probe. Acta Biochim Biophys Sin 2002 34 11-5... 

Various chemiluminescent compounds (e.g. luminol) have been studied in order to find suitable and specific probes for detection of ROS. Recently Cypridina luciferin analogues CLA and MCLA are thought to emit light only when reacting with superoxide anion and singlet oxygen. ... 

Spectra of the superoxide-induced chemiluminescence of probes A-1, A-2, and B exhibited their luminescence maximum only at around 515 - 527 ran, which was due to luminescence from the fluorescein moiety, along with the absence of blue luminescence due to the MCLA moiety. These results clearly indicated that the superoxide-induced chemiluminescence of A-1, A-2, and B were generated fi-om the... 

This study showed that probe B, in which MCLA and fluorescein molecules were bound at the secondary and primary faces of y-cyclodextrin, respectively, was successfully prepared. Subsequent characterization of probe B demonstrated that it can generate green light, possesses high sensitivity to superoxide anions, and exhibits high chemiluminescence intensity, in comparison to FCLA. 

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