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The 3D reconstitution was done using the Amira 3D v4.1.1 Imaging program.
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Reconstitution is done by soaking in hot, but not boiling, water for about twenty minutes; the water used is infused with the mushroom aroma and it too can be used in subsequent cooking.
Deleting a wild-type protein or replacing it with a purified mutant protein during reconstitution, as was done here, provides the same flexibility in a mammalian system that is achieved genetically in yeast.
The reconstitution of HIE was done in methanol to prepare stock solution of 100 mg/ml for photochemical and in vitro analysis and in 1% DMSO for in vivo dosage preparation.
Reconstitution of samples was done one at a time due to the PTC amino acid sensitivity to light and ambient temperature.
The precipitate was removed by centrifugation for 20 min at 18000 g at 4°C, the supernatant applied to an NAP 10 column (GE Healthcare), and sAC-HD eluted with buffer A. Reconstitution of holo protein was done by titration in increments of 5 μM using a 1 mM haemin stock solution.
Vaccine reconstitution by the unblinded nurse was done in a secure room, and then the individual dose carried to the participant's room on a tray covered by an opaque cloth.
Reconstitution of the Ripk1 −/− MEFs was done by lentiviral transduction.
Chest roentgenogram was done at baseline, at six months post ATT, and at the time of Immune Reconstitution Inflammatory syndrome (IRIS), if applicable.
Reconstitution and assays were done the same day.
Reconstitution and quality control of the final solution were done according to the procedure described in the Summary of Product Characteristics (SmPC).
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