Fig. 10: Pulldown of the native PfKelch13 protein from P. falciparum total protein extracts using hemin-agarose beads.

3D7 parasites were isolated by saponin treatment from asynchronized culture and proteins were solubilized with RIPA buffer. Protein extracts were incubated with hemin-agarose or glutathione (GSH)-agarose beads. After extensive washing, bound proteins were eluted with Laemmli buffer, separated by SDS-PAGE, and blotted against PfKelch13 antibodies. PfKelch13 antibodies were custom-generated using a PfKelch13 peptide51. Molecular weight standards (in kDa) are as indicated.