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To analyze protein production, soluble proteins in the spent culture medium were separated on a SDS-polyacrylamide 12.5% gel (ATTO, Tokyo, Japan) and stained with Bio-Safe Coomassie Stain (Bio-Rad, Hercules, CA).
The flocs and the growth medium were separated after flocculation.
Cells and medium were separated by centrifugation at 12.000 ×g for 10 min at room temperature.
Proteins from cell lysates and conditioned medium were separated by SDS-PAGE.
After incubation at 26°C for 6 days, the mycelia and culture medium were separated by centrifugation at 96 ×g.
hESC-conditioned medium was incubated with Heparin-Agarose Beads for 2 hours shaking at 4C. Beads and all medium were separated by centrifugation.
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There is a reason RPGs and other "deep" games work well using keyboards and, to a degree, controllers: the medium is separated from the message of the game.
After the incubation period, the culture medium was separated from the cultured cells and subjected to a LDH assay that was carried out following the manufacturer's instructions.
Firstly, the conditioned medium was separated to two parts by ultra filter.
One hundred micrograms of protein from the concentrated conditioned medium was separated by SDS-PAGE, dintoed intenten horizontal slices, and subject to trypsinization and LC-MS/MS (see Figure S1 and Materials and Methods).
A total of 1 μg of protein from each conditioned medium was separated on 10% polyacrylamide gels containing 0.2% gelatin.
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