Cy2 conjugated donkey anti mouse igg

To detect SARS-CoV-2 seroprevalence in the plasma samples, an immunofluorescence assay (IFA) against the SARS-CoV-2 spike and nucleocapsid proteins was used as previously described [18 (link)]. Briefly, transfected HEK-293T cells (ATCC, Manassas, VA, USA) expressing either the spike or nucleocapsid proteins of SARS-CoV-2 (Addgene, Watertown, MA, USA and Sino Biological, Chesterbrook, PA, USA, respectively) were fixed and seeded onto 12-well polytetrafluoroethylene (PTFE)-printed slides (Electron Microscopy Sciences, Hatfield, PA, USA). The plasma samples were diluted 1:20 in 1× PBS with 0.1% of Tween 20, and 15 µL of the diluted plasma sample was added to each corresponding well on the slide and incubated for 1 h at 37 °C. After washing, the slides were incubated with secondary mouse monoclonal anti-human IgG antibody (ATCC, Manassas, VA, USA), followed by the removal of unbound antibodies and incubation with tertiary CY2-conjugated donkey anti-mouse IgG (Jackson ImmunoResearch Laboratories, West Grove, PA, USA). The cells were then counterstained with Evans blue solution. The stained IFA was evaluated by three independent readers using a Nikon Eclipse 50i fluorescence microscope to determine positive or negative signals on the slide, and only concordant results from at least two independent readers were reported as the outcome.

Rabbit polyclonal anti-Arl2 (Cat#: 10232-1-AP) and rabbit anti-Arf5 (Cat#: 20227-1-AP) were purchased from Proteintech Inc. (Chicago, IL, USA). Mouse monoclonal anti-α-tubulin antibody (Cat#: sc-8035) antibody was purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). Human anti-centromere CREST antibody (Cat#: 15-234) was purchased from Antibodies Incorporated (Davis, CA, USA). Cy2-conjugated donkey anti-mouse IgG (Cat#: 715-225-150), Cy2-conjugated donkey anti-rabbit IgG (Cat#: 711-225-152) and Cy3-conjugated donkey anti-human IgG (Cat#: 711-225-152) were purchased from Jackson ImmunoResearch Laboratory (West Grove, PA, USA).