Quant ittm picogreen dsdna reagent and kits

DNA in the plasma of human and mice was quantified according to the manufacturer’s instructions using Quant-iT^TM PicoGreen ® dsDNA Reagent and Kits (Invitrogen, MA, USA). The fluorescence intensity reflects the amount of DNA, which was measured at excitation and emission wavelengths of 480 nm and 520 nm, respectively, in a microplate reader (Thermo Fisher Scientific, MA, USA).

Ds DNA in the plasma of mice was detected using Quant-iT^TM PicoGreen ^® dsDNA Reagent and Kits (Invitrogen, Carlsbad, CA, USA). As described, the obtained plasma and diluted standard DNA were added to a Picogreen working solution and incubated for 2 to 5 min at room temperature, protected from light. Fluorescence intensity was measured by a fluorescence microplate reader at a 480 nm excitation wavelength and 530 nm emission wavelength. The amount of DNA was determined from the standard curve.
The concentration of MPO-DNA complexes in the plasma of mice was determined using a capture ELISA kit, as previously reported. An anti-MPO antibody (22225-1-AP, Proteintech, Wuhan, China) was coated overnight at 4 °C for the antibody capture. After blocking with 1% BSA, the plasma was added to the wells with peroxidase-labelled anti-DNA antibody and incubated for 2 h at room temperature. Next, the plate was washed, and peroxidase substrate was added for 40 min of incubation at 37 °C, protected from light. Optical density was determined at 405 nm.