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Prior to LC-MS analysis, the buffer was exchange to 10 mM ammonium acetate.
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b The T-tube was exchanged to a locking pigtail drain catheter that drained externally.
The buffer was exchanged to 10mM sodium-phosphate (pH 7.0) and the SPRR proteins were stored at −80°C.
The solvent was exchanged to hexane and the fat content was determined gravimetrically.
The buffer was exchanged to 100 mM AmAc for MS analysis.
Medium was exchanged to DMEM with GlutaMax™ without FCS after 4 hours.
The medium was exchanged to the fresh serum-containing medium after 4 h.
After 3 weeks the cast was exchanged to one in neutral position, for another 3 weeks.
Four hr later, the medium was exchanged to neuronal maintenance medium.
After 20 min, the medium was exchanged to remove most of the contaminating astrocytes.
On days 0, 3, and 7 half of the media was exchanged to minimize the inflammatory burden.
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