H2O2 production was determined by a Luminol-based assay. Four mm diameter disc leaves from 4-5 week-old Arabidopsis thaliana plants (n = 8) were collected in 96-wells white plates (Thermo Scientific) and incubated overnight in 100 μl of ROS Buffer (100 μg/ml peroxidase, Sigma; and 100 nM Luminol, Sigma). Luminescence was measured as RLU (relative light units) every minute over 40 minutes after induction with 1 μM flg22 or 50 μM chitohexaose (CHI6) in a Varioskan LUX luminometer (Thermo Scientific), as described in Torres et al. (2013) (link).
Varioskan lux luminometer
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Varioskan LUX luminometer is a multi-mode microplate reader designed for versatile detection of luminescent, fluorescent, and absorbance-based assays. It features a high-performance optical system and provides rapid, sensitive, and flexible read capabilities for a wide range of sample types and plate formats.
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Lab products found in correlation
2 protocols using varioskan lux luminometer
1
Measuring H2O2 Production in Arabidopsis
2
Luminol-Based Oxidative Burst Assay
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Twelve leaves from 10–day–old rice or 2–week–old wheat seedlings were used to generate leaf discs of 0.25 cm2 using a hole punch and were incubated in water overnight in a 96–well plate to eliminate the effects of wounding. The next morning water was replaced by 100 μL reaction solution containing 200 μm luminol L–012 (Wako, Japan) and 1 μg horseradish peroxidase (Sigma, St. Louis, MO, USA) supplemented with 10 μm OsPep plus 0.05% silwet L–77 or silwet L–77 only (mock). The luminescence was recorded by a Varioskan Lux luminometer (Thermo, Waltham, MA, USA) for a period of 30 min.
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