L 012 sodium salt

L 012 sodium salt is a chemiluminescent probe [1].

L 012 sodium salt is a new chemiluminescent probe. In activated EoL-1 cells which generated reactive oxygen species (ROS), treatment with L-012 generated a significant chemiluminescence. While, superoxide dismutase (100-300 U/ml) abolished the chemiluminescence, which suggested that the main ROS detected was O2- [1]. In cultured endothelial cells, phorbol 12-myristate 13-acetate (PMA, 1 μM) caused an 2.8-fold increase of L-012 chemiluminescence. Vanadate (100 μM) increased the chemiluminescence up to 17-fold. These results suggested that the increase of superoxide produced by NAD(P)H oxidase [2].

In human oral cavity and blood and rat peritoneal cavity, L-012 reacted with ROS generated by activated neutrophils and generated strong chemiluminescence, which was higher than that of MCLA [3]. In nonobese diabetic mice and K8 knockout (K8(-/-)) mice, treatment with dextran sulphate sodium generated an increased L-012 chemiluminescence which detected reactive oxygen and nitrogen species (ROS and RNS) [4].

References:
[1].  Nishinaka Y, Aramaki Y, Yoshida H, et al. A new sensitive chemiluminescence probe, L-012, for measuring the production of superoxide anion by cells. Biochem Biophys Res Commun, 1993, 193(2): 554-559.
[2].  Sohn HY, Gloe T, Keller M, et al. Sensitive superoxide detection in vascular cells by the new chemiluminescence dye L-012. J Vasc Res, 1999, 36(6): 456-464.
[3].  Imada I, Sato EF, Miyamoto M, et al. Analysis of reactive oxygen species generated by neutrophils using a chemiluminescence probe L-012. Anal Biochem. 1999, 271(1): 53-58.
[4].  Asghar MN, Emani R, Alam C, et al. In vivo imaging of reactive oxygen and nitrogen species in murine colitis. Inflamm Bowel Dis, 2014, 20(8): 1435-1447.

Evaluation of lophine derivatives as L-012 (luminol analog)-dependent chemiluminescence enhancers for measuring horseradish peroxidase and H2O2.[Pubmed:23630098]

Luminescence. 2014 Mar;29(2):118-21.

8-Amino-5-chloro-7-phenylpyrido[3,4-d]pyridazine-1,4(2H,3H)dione (L-012) was recently synthesized as a new chemiluminescence (CL) probe; the light intensity and the sensitivity of L-012 are higher than those of other CL probes such as luminol. Previously, our group developed four lophine-based CL enhancers of the horseradish peroxidase (HRP)-catalyzed CL oxidation of luminol, namely 2-(4-hydroxyphenyl)-4,5-diphenylimidazole (HDI), 2-(4-hydroxyphenyl)-4,5-di(2-pyridyl)imidazole (HPI), 4-(4,5-diphenyl-1H-imidazol-2-yl)phenylboronic acid (DPA), and 4-[4,5-di(2-pyridyl)-1H-imidazol-2-yl]phenylboronic acid (DPPA), and showed that DPPA was suitable for the photographic detection of HRP. In this study, we replaced luminol with L-012 and evaluated these as L-012-dependent CL enhancers. In addition, to detect HRP and/or H2O2 with higher sensitivity, each detection condition for the L-012-HRP-H2O2 enhanced CL was optimized. All the derivatives enhanced the L-012-dependent CL as well as luminol CL; HPI generated the highest enhanced luminescence. Under optimized conditions for HRP detection, the detection limit of HRP was 0.08 fmol. By contrast, the detection limit of HRP with the enhanced L-012-dependent CL using 4-iodophenol, which is a common enhancer of luminol CL, was 1.1 fmol. With regard to H2O2 detection, the detection limits for enhanced CL with HPI and 4-iodophenol were 0.29 and 1.5 pmol, respectively. Therefore, it is demonstrated that HPI is the most superior L-012-dependent CL enhancer.

Noninvasive assessment of localized inflammatory responses.[Pubmed:22080048]

Free Radic Biol Med. 2012 Jan 1;52(1):218-26.

Inflammatory diseases are associated with the accumulation of activated inflammatory cells, particularly polymorphonuclear neutrophils (PMNs), which release reactive oxygen species (ROS) to eradicate foreign bodies and microorganisms. To assess the location and extent of localized inflammatory responses, L-012, a highly sensitive chemiluminescent probe, was employed to noninvasively monitor the production of ROS. We found that L-012-associated chemiluminescence imaging can be used to identify and to quantify the extent of inflammatory responses. Furthermore, regardless of differences among animal models, there is a good linear relationship between chemiluminescence intensity and PMN numbers surrounding inflamed tissue. Depletion of PMNs substantially diminished L-012-associated chemiluminescence in vivo. Finally, L-012-associated chemiluminescence imaging was found to be a powerful tool for assessing implant-mediated inflammatory responses by measuring chemiluminescence intensity at the implantation sites. These results support the use of L-012 for monitoring the kinetics of inflammatory responses in vivo via the detection and quantification of ROS production.

In vivo imaging of reactive oxygen and nitrogen species in inflammation using the luminescent probe L-012.[Pubmed:19539751]

Free Radic Biol Med. 2009 Sep 15;47(6):760-6.

Production of reactive oxygen and nitrogen species (ROS/RNS) is an important part of the inflammatory response, but prolonged elevated levels of ROS/RNS as under chronic inflammation can contribute to the development of disease. Monitoring ROS/RNS in living animals is challenging due to the rapid turnover of ROS/RNS and the limited sensitivity and specificity of ROS/RNS probes. We have explored the use of the chemiluminescent probe L-012 for noninvasive imaging of ROS/RNS production during inflammation in living mice. Various inflammatory conditions were induced, and L-012-dependent luminescence was recorded with an ultrasensitive CCD camera. Strong luminescent signals were observed from different regions of the body corresponding to inflammation. The signal was reduced by administration of the SOD mimetic tempol, the NADPH oxidase inhibitor apocynin, and the inhibitor of nitric oxide synthesis L-NAME, signifying the requirement for the presence of ROS/RNS. Additionally, the L-012 signal was abolished in mice with a mutation in the Ncf1 gene, encoding a protein in the NADPH oxidase complex 2, which generates ROS/RNS during inflammation. In conclusion, L-012 is well distributed in the mouse body and mediates a strong ROS/RNS-dependent luminescent signal in vivo and is useful for monitoring the development and regulation of inflammation in living organisms.