Parasite Isolation and Lysis Protocol

P. falciparum cultures were prepared as described previously (21 (link), 29 (link)). Erythrocytes were then lysed with 0.0125% saponin (Sigma, sapogenin content ≥ 10%) in cold phosphate-buffered saline (PBS) with an EDTA-free protease inhibitor mixture (Roche or Pierce). The released P. falciparum parasites were then washed in cold PBS with EDTA-free protease inhibitor mixture. The washed cell pellets were flash frozen in liquid nitrogen and stored at −80 °C.

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Ho C.M., Jih J., Lai M., Li X., Goldberg D.E., Beck J.R, & Zhou Z.H. (2021). Native structure of the RhopH complex, a key determinant of malaria parasite nutrient acquisition. Proceedings of the National Academy of Sciences of the United States of America, 118(35), e2100514118.

Publication 2021

saponin sapogenin

Cell Cold Edta Erythrocytes Frozen Nitrogen Parasites Pellets Phosphate Protease inhibitor Saline Saponin

Corresponding Organization : California NanoSystems Institute

Other organizations : Washington University in St. Louis, Iowa State University

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Variable analysis

independent variables

Concentration of saponin (0.0125%)

dependent variables

Lysis of erythrocytes
Release of P. falciparum parasites

control variables

Phosphate-buffered saline (PBS)
EDTA-free protease inhibitor mixture (Roche or Pierce)
Low temperature (cold PBS, flash freezing in liquid nitrogen, storage at -80°C)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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