Alexa 488 conjugated goat anti mouse igg h l

Cell monolayers were fixed with ice-cold methanol and then air-dry. For the detection of N protein expression, cells were incubated with a mAb (4A5) at 37°C for 1 h. After five washes, the cell monolayers were stained with Alexa 488-conjugated goat anti-mouse IgG(H + L; Jackson ImmunoResearch Inc., West Grove, PA, United States). Cell nuclei were counterstained with 4′,6-diamidino-2-phenylindole (DAPI) solution (Solarbio Life Sciences, Beijing, China) for 5 min at room temperature. After extensive washes with 1×PBS, fluorescent images were captured with the IX73 epifluorescence microscope (Olympus).

A thin smear of schizont-rich culture of P. berghei was used for immunocytochemistry. Immunocytochemistry was carried out as previously described with some modifications^{29 (link)}. Briefly, parasites on the slides were fixed in ice-cold acetone for 10 min and then blocked in 3% BSA-PBS for 30 min. The fixed parasites were incubated with primary antibodies overnight at 4 °C, washed, and then incubated with secondary antibodies for 60 min at room temperature. The slides were mounted with ProLong Diamond Antifade Mountant with DAPI (Thermo Fisher Scientific) and analyzed under a standard fluorescence microscope (Olympus). Mouse anti-FLAG M2 antibody (1:1000; Sigma, F1804-200UG) was used for the detection of FLAG-tagged Cas9. Alexa488-conjugated goat anti-mouse IgG (H + L) (1:200, Jackson #A-11029) was used as a secondary antibody.