The lyophilized extracts (400 µg protein) were dissolved in rehydration solution (8 M urea, 2 M thiourea, 19.4 mM DTT, 1% CHAPS, 0.5% carrier ampholytes (Pharmalyte, pH 3 10, Amersham Bioscience) and traces of BPB) and were separated in the first dimension on Immobilized Dry Strip Gels, pH 4 7/180 mm (Amersham Bioscience).
Overlapping peaks were separated using traces of single ions.
Expression traces corresponding to these two classes were separated throughout the experiment.
FAMEs were separated using a Trace 1300 gas chromatograph (GC) (Thermo-Scientific, USA) on a 30 m FAMEWAX column (Restek #12497, Bellefonte, USA) and detected using a TSQ 8000 triple quadrupole detector (Thermo-Scientific, Waltham, USA) operated in full scan mode (50 – 550 m/z).
The tables of image and trace files, such as bc_file_img and bc_file_trace, were separated to speed up and manage attachments easily.
Unlike the trace metals and REEs, Ca was accumulated in coarser particles that were separated by underflow fractions.
Both the layers were separated and oils were collected in clear vials, after removing water traces using anhydrous sodium sulfate, and they were stored at 4 °C until needed.
Purified CD4+ T cells from PBMCs were separated using the CD4+ T-cell Isolation Kit II (Miltenyi Biotech) and stained with Cell Trace Violet (Invitrogen).
Alternatively, purified CD4+ T cells from tonsils were separated using the CD4+ T-cell Isolation Kit II (Miltenyi Biotech) and stained with Cell Trace Violet (Invitrogen).
Let sit until all traces of food are separated from the sieve.
