The
luminol and lucigenin levels, bcl-2/bax ratio, [Na.sup.+]/[K.sup.+]-ATPase, caspase-3, and caspase-9 activities of the frozen samples were examined once thawed at the room temperature.
Antioxidant Capacity Assay by
Luminol Chemiluminescence.
It is based on CR1/CR3 receptor activation of neutrophils in the whole blood, which produce oxidants that react with
luminol [19].
The ROS levels in the rat retina were analyzed using
luminol (5-amino-2,3-dihydro-1,4-phthalazinedione, Sigma) and lucigenin (bis-N-methylacridiniumnitrate, Sigma).
Briefly, using the chemiluminescence technique to measure the antioxidant capacity of biological fluids, the light emission occurs when the chemiluminescent substrate (i.e.,
luminol) is oxidized by [H.sub.2][O.sub.2] in a reaction catalyzed by "horseradish peroxidase." The stabilities and intensities of the light emitted are high due to the addition of p-iodine phenol.
Katsu, "Porphyrin-induced photogeneration of hydrogen peroxide determined using the
luminol chemiluminescence method in aqueous solution: a structure-activity relationship study related to the aggregation of porphyrin," Analytical Sciences, vol.
Briefly, the macrophages were adhered to round glass coverslips (9 mm) at concentration of 1 x [10.sup.5] cells per coverslip and then placed in cuvettes containing 1 ml of Hank's solution (
luminol 0.8 x [10.sup.-4]M and HRP at 4U).
The most common method for detecting blood stains is to spray a crime scene with
luminol, a chemical that reacts with iron in blood to emit a blue glow that can be seen in the dark; the use of
luminol has its disadvantages.
Camptothecin (98% purity),
luminol, f-MLP (formyl-met-leuphenylalanine), SAAVNA (N-succinyl-alanine-alanine-valinine-pnitroanilide), cytochalasin B, TMB (3,3',5, 5'-tetramethylbenzidine) liquid substrate system, Hanks' balanced salt solution (HSSB), L-glutamine, fetal bovine serum (FBS), 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) and RPMI1640 medium were purchased from Sigma-Aldrich GmbH (Steinheim, Germany).