Freshly egressed tachyzoites were 3.0 µm-filter purified, spun down and resuspended in Hank's balanced salt solution supplemented with 100 mM HEPES (HBSS-H), 20 mM EGTA at ∼5×106 parasites/ml. All the assays were done in HBSS not containing Mg2+ or Ca2+, unless indicated. Parasites were layered onto poly-L-lysine-coated coverslips and allowed to adhere for 5–15 min at RT then incubated for 20 min at 37°C, in the presence or absence of 2 mM ionomycin (Santa Cruz Biotechnology, 56092-82-1 Ionomycin-3592). After incubation, parasites were fixed as above followed visualisation of gliding trails using α-SAG1 (Table S2 ) with no permeabilisation. Images were acquired using a DeltaVision Core microscope (AppliedPrecision, GE Healthcare) as described (Ovciarikova et al., 2017 (link)). Image brightness was adjusted using FIJI software to allow for clear visualisation of the trails, which were then traced with a wand tool, and the length of the trail was measured.
56092 82 1 ionomycin 3592
Manufactured by Santa Cruz Biotechnology
56092-82-1 Ionomycin-3592 is a chemical compound used in laboratory research applications. It functions as a calcium ionophore, facilitating the movement of calcium ions across cell membranes. This product is intended for use in scientific investigations, but its specific applications should be determined by the researcher.
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2 protocols using 56092 82 1 ionomycin 3592
1
Quantifying Toxoplasma Gliding Motility
2
Quantifying Tachyzoite Gliding Motility
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Freshly egressed tachyzoites were 3.0 µm-filter purified, spun down and resuspended in Hank's balanced salt solution supplemented with 100 mM HEPES (HBSS-H), 20mM EGTA at around 5 x 10 6 parasites/mL. All the assays were done in HBSS not containing Mg or Ca, unless indicated. Parasites were layered onto poly-L-lysine-coated coverslips and allowed to adhere for 5 -15 min at RT then incubated for 20 min at 37 °C, in the presence or absence of 2 mM ionomycin (Santa Cruz Biotechnology, 56092-82-1 Ionomycin-3592). After incubation, parasites were fixed as above followed visualization of gliding trails using the α-SAG1 (Table S2) with no permeabilisation. Images were acquired using a Delta Vision microscope as described (Ovciarikova et al., 2017) (link). Image brightness was adjusted using FIJI software to allow for clear visualisation of the trails which were then traced with a wand tool and length of the trail was measured.
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