Goat anti rabbit hrp conjugate

Nuclear and cytoplasmic extracts from parasites were prepared as previously described (Voss et al. 2002 ) using 5–7% of the synchronized parasites. The proteins from the nuclear and cytoplasmic fractions were run on a gel (mini PROTEAN precast TGX gels) and then transferred to a nitrocellulose membrane. The membrane was then blocked using 5% w/v non-fat milk powder in 1X PBS-0.05% v/v Tween for 45 minutes at room temperature. The membrane was then incubated overnight at 4°C with an anti-GFP (ROCHE Anti-GFP, from mouse IgG1K, Millipore Sigma) (1:1000), anti-histone (Anti-Histone H3 antibody, AbCam) (1:3000), or anti-aldolase (Anti-Plasmodium aldolase antibody (HRP), AbCam) (1:1000) primary antibody in the blocking buffer. After incubation, the membrane was washed three times with 1X PBS-0.05% v/v Tween and was incubated with the appropriate secondary antibodies, then washed. The secondary antibodies used were as follows: peroxidase-conjugated goat anti-rat HRP conjugate (Millipore) / goat anti-rabbit HRP conjugate (Millipore) / goat anti-mouse HRP conjugate (Pierce) was used (1:3000). Bound antibodies were then visualized on a film after enhancing the signal with ECL (Pierce) as the substrate.