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Previously reported conditions for generating trypsin-α2M vary greatly [2,5,20]; therefore, in this study we used an optimized method to obtain preparations of trypsin-α2M that were completely activated but not degraded (Supplementary Fig. 1).
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The obtained preparations were >95% pure and viable.
The presence of phospholipids in the buffers was an absolute requirement for obtaining preparations of enzyme that were capable of ATP synthesis after reconstitution into phospholipid vesicles.
Thus, we obtained preparations of amastigotes associated with host phagolysosomal components (LIMP1 positive), mimicking parasite-containing host cell extrusions, which display or are devoid of LAMP1/LAMP2 proteins (after amastigote isolation from MEF-WT and MEF-LAMP1/2KO respectively).
Thus, obtained preparation was used for further analysis.
The obtained preparation was stained by the Papanicolaou method for cytological examination.
The obtained preparation was ultradiafiltered against PBS, using YM10 membrane (Amicon Division, W.R. Grace, Beverly, MA).
The antioxidant and antibacterial properties of such obtained preparation were analysed, in a purpose, to find the best source to generate antimicrobial preparation based on flax.
These values were close to those recorded in the previous report [ 4] when lettuce bearing the S antigen was lyophilised without any protective excipients and the obtained preparation showed good stability over one-year storage at room temperature.
Isolated mitochondria were sonicated on ice to obtain mtDAMP preparations.
It was technically impossible to obtain microvessel preparations of adequate purity from rat brain tissue at ages earlier than 9 days [18].
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