Quantification of NET-bound MPO

Briefly, plasma or supernatant from neutrophils culture was incubated overnight in a plate precoated with anti-MPO antibody (Thermo Fisher Scientific; cat. PA5-16672). The DNA bound to MPO was quantified using the Quant-iT PicoGreen kit (Invitrogen; cat. P11496), as described (Colón et al., 2019 (link); Czaikoski et al., 2016 (link)).

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Veras F.P., Pontelli M.C., Silva C.M., Toller-Kawahisa J.E., de Lima M., Nascimento D.C., Schneider A.H., Caetité D., Tavares L.A., Paiva I.M., Rosales R., Colón D., Martins R., Castro I.A., Almeida G.M., Lopes M.I., Benatti M.N., Bonjorno L.P., Giannini M.C., Luppino-Assad R., Almeida S.L., Vilar F., Santana R., Bollela V.R., Auxiliadora-Martins M., Borges M., Miranda C.H., Pazin-Filho A., da Silva L.L., Cunha L., Zamboni D.S., Dal-Pizzol F., Leiria L.O., Siyuan L., Batah S., Fabro A., Mauad T., Dolhnikoff M., Duarte-Neto A., Saldiva P., Cunha T.M., Alves-Filho J.C., Arruda E., Louzada-Junior P., Oliveira R.D, & Cunha F.Q. (2020). SARS-CoV-2–triggered neutrophil extracellular traps mediate COVID-19 pathology. The Journal of Experimental Medicine, 217(12), e20201129.

Publication 2020

anti-MPO antibody Quant-iT PicoGreen kit

Anti antibody Neutrophils Picogreen Plasma

Corresponding Organization : Universidade de São Paulo

Other organizations : Universidade do Sul de Santa Catarina

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Variable analysis

independent variables

Incubation of plasma or supernatant from neutrophils culture overnight in a plate precoated with anti-MPO antibody

dependent variables

DNA bound to MPO

control variables

The use of Quant-iT PicoGreen kit (Invitrogen; cat. P11496) to quantify the DNA bound to MPO, as described in the referenced studies (Colón et al., 2019; Czaikoski et al., 2016)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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