The supernatant was collected, concentrated, filtered and maintained at 4°C for dissociated and explant culture treatment.
Fractions were concentrated, filtered (0.45 µm), and further purified to homogeneity using a Superdex 75 size exclusion column (GE Healthcare) equilibrated with 20 mM HEPES (pH 7.5), 100 mM NaCl, 1 mM EDTA and 0.5 mM DTT.
All dust samples were extracted with acetone:hexane (1 1, vol/vol) using sonication, and then concentrated, filtered, and reconstituted in dimethyl sulfoxide (DMSO).
All conditioned medium was sterile filtered through a 22-μm filter to remove cellular debris and concentrated using a 3000 kDa centrifugal concentrating filter (Amicon, Billerica, MA, USA) to give 500 μl (i.e., concentrated ×30).
All ore coming out of the mines is low-grade, with iron content of around 35%. Pelletizing that ore a laborious process of grinding, separating, concentrating, filtering, drying and heating bumps that content up to 65% or so.
Moreover, the method concentrates the filter adaptation to the filter which is most misadjusted, which is a typically desired feature.
The KI-soluble material was then concentrated and filtered using a Centricon YM-10 (Millipore) device.
Cells were then concentrated by filtering through 10 μM sieve (Sefar American).
After that, supernatant was collected, filtered, concentrated 2X using Centriprep® filter device (Millipore) and stored at −80°C.
The reaction mixture was filtered, concentrated, dissolved in dichloromethane, filtered, and concentrated again to yield the desired product as an off-white solid (72.6 mg, 43% yield).
