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It is a heavy standard but one that is not unreasonable.
She was in her N.Y.P.D. uniform complete with all the equipment — gun, handcuffs, radio, etc. — and she wore the heavy standard police outer coat.
NR content in conditioned media was measured using matrix-assisted laser desorption ionization (MALDI) MS. A heavy standard of NR was synthesized from hydrolysis of cyanopyridine in 18O water to produce 18O Nam [27], followed by chemical synthesis of heavy NR from heavy Nam [28].
For absolute quantification of NRX-beta isoforms full-length NRX-beta fused to GFP was used as heavy standard.
The heavy standard in turn served to deduce absolute amounts of the endogenous proteins in the brain extracts using target protein-specific peptides.
Tryptic peptide mixtures including heavy standard peptides were then analysed by SRM using nanoflow HPLC (Easy nLC, Thermo Scientific, Ulm, Germany) coupled to a triple quadrupole mass spectrometer as mass analyser (TSQ Quantum Discovery Max, Thermo Scientific, Ulm, Germany).
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These "heavy standards", as they are called, have to be anchored underground and take much longer than a smaller tree to establish themselves safely.
In addition, the amount of NRX-alpha was calculated in samples measured with beta heavy standards.
Quantification was performed with NRX-alpha and beta heavy standards spiked into the brain samples.
For absolute quantification of NRX-alpha proteins, the neurexin-repeat 3 containing laminin-G-like domains 4 and 6 (LG4-6) fused to GFP was used as heavy standards.
The peptides sequences and transitions for monitoring peptides and heavy standards are listed in Table 1 and Supporting Information Table S1.
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