Immunofluorescence Assay for PfAP2-EXP2 Localization

The immunofluorescence assay was performed to detect the localization of PfAP2-EXP2 as described previously (Jing et al., 2018 (link)). Parasites were harvested, released, fixed by 4% paraformaldehyde (Electron Microscopy Sciences) at room temperature for 10 min, and washed by PBS. Prepared samples were incubated with the primary antibodies against ty1 (Sigma, SAB4800032) or GFP (Abcam, ab290) at 1:500 to 1:1,000, followed by the secondary antibodies AlexaFluor 488 goat anti-mouse IgG (ThermoFisher Scientific, A11029) or AlexaFluor 568 goat anti-rabbit IgG (ThermoFisher Scientific, A11036) at 1:500. Preparations were visualized with a Nikon A1R microscope at 60-100 × magnification and images were acquired with NIS Elements software and processed using Adobe Photoshop.

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Shang X., Wang C., Shen L., Sheng F., He X., Wang F., Fan Y., He X, & Jiang M. (2022). PfAP2-EXP2, an Essential Transcription Factor for the Intraerythrocytic Development of Plasmodium falciparum. Frontiers in Cell and Developmental Biology, 9, 782293.

Publication 2021

4% paraformaldehyde SAB4800032 ab290 AlexaFluor 488 goat anti-mouse IgG AlexaFluor 568 goat anti-rabbit IgG A1R microscope

Anti igg Antibodies Electron microscopy Goat Immunofluorescence assay Microscope Mouse Paraformaldehyde Parasites Rabbit

Corresponding Organization : Jiangsu Institute of Parasitic Diseases

Other organizations : Central South University

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Variable analysis

independent variables

Primary antibodies against ty1 (Sigma, SAB4800032) or GFP (Abcam, ab290) at 1:500 to 1:1,000

dependent variables

Localization of PfAP2-EXP2

control variables

Parasite samples were harvested, released, fixed by 4% paraformaldehyde at room temperature for 10 min, and washed by PBS
Prepared samples were incubated with the primary antibodies
Secondary antibodies AlexaFluor 488 goat anti-mouse IgG (ThermoFisher Scientific, A11029) or AlexaFluor 568 goat anti-rabbit IgG (ThermoFisher Scientific, A11036) at 1:500
Preparations were visualized with a Nikon A1R microscope at 60-100 × magnification and images were acquired with NIS Elements software and processed using Adobe Photoshop

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