2.13. Flow cytometry analysis of ZIKV infection

Mônika Bezerra dos Santos Oliveira; Iara Barros Valentim; Tauane dos Santos Rocha; Jaqueline Correia Santos; Keyla Silva Nobre Pires; Eloiza Lopes Lira Tanabe; Karen Steponavicius Cruz Borbely; Alexandre Urban Borbely; Marília Oliveira Fonseca Goulart

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2.13. Flow cytometry analysis of ZIKV infection

MO Mônika Bezerra dos Santos Oliveira

IV Iara Barros Valentim

TR Tauane dos Santos Rocha

JS Jaqueline Correia Santos

KP Keyla Silva Nobre Pires

ET Eloiza Lopes Lira Tanabe

KB Karen Steponavicius Cruz Borbely

AB Alexandre Urban Borbely

MG Marília Oliveira Fonseca Goulart

This method is extracted from research article: Ind Crops Prod, Apr 2020

Schinus terebenthifolius Raddi extracts: from sunscreen activity toward protection of the placenta to Zika virus infection, new uses for a well-known medicinal plant

DOI: 10.1016/j.indcrop.2020.112503

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To access the percentage of cells infected by ZIKV, cells were detached with 2.5% trypsin-EDTA and fixation and permeabilization were performed using commercial flow cytometry kits (e-Bioscience, San Diego, CA, EUA). A monoclonal primary mouse antibody anti-ZIKV NS1 (1:100, E107, MA5-24585, Invitrogen) was incubated with Zenon Alexa 488 conjugated secondary anti-mouse antibody (1:250; Thermo Fisher Scientific) for 5 min to form an immunocomplex with further addition of Zenon Blocking Reagent for further 5 min. The immunocomplex was later incubated with cells for 1 h at 37 °C. The percentage of ZIKV infected cells was analysed with FACS CantoTM® II flow cytometer (BD Biosciences, San Jose, CA, USA), using the software FlowJo (TreeStar, Ashland, OR, USA).

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