HPLC/analytical grade reagents were used throughout. 1,1-carbonydiimidiazole, triethylamine, iodomethane, iodomethane-d3, iodoethane, iodoethane-d5, iodopropane, formic acid, citric acid, methanol, disodium hydrogen phosphate, chloroform, ammonium formate, acetonitrile, 1-butanol, and analytical standards for saturated and unsaturated fatty acids were purchased from Sigma-Aldrich. All lipid extractions were performed in 2 mL polypropylene LoBind safe-lock tubes (Eppendorf).
Cell culture reagents were purchased from Life Technologies. Mouse anti-STXBP1 antibody was purchased from BD Biosciences. pCMV-STXBP1-emGFP, pCMV-STXBP1, NPY-hPLAP, and NPY-mCherry were prepared as previously described (Arunachalam et al, 2008 (link); Martin et al, 2013 (link); Tomatis et al, 2013 (link)). STXBP1Δ317-333 and STXBP1F115E mutants were made using the quick-change lightning site-directed mutagenesis kit (Strategene, USA) and the mutational primer
5′-GACTTTTCCTCTAGCAAGAGGATGATGCCCCAGTACCAGAAGGAGC-3′, as previously described (Martin et al, 2013 (link)). All constructs were sequenced at The Australian Genome Research Facility, located at The University of Queensland.
Cell culture reagents were purchased from Life Technologies. Mouse anti-STXBP1 antibody was purchased from BD Biosciences. pCMV-STXBP1-emGFP, pCMV-STXBP1, NPY-hPLAP, and NPY-mCherry were prepared as previously described (Arunachalam et al, 2008 (link); Martin et al, 2013 (link); Tomatis et al, 2013 (link)). STXBP1Δ317-333 and STXBP1F115E mutants were made using the quick-change lightning site-directed mutagenesis kit (Strategene, USA) and the mutational primer
5′-GACTTTTCCTCTAGCAAGAGGATGATGCCCCAGTACCAGAAGGAGC-3′, as previously described (Martin et al, 2013 (link)). All constructs were sequenced at The Australian Genome Research Facility, located at The University of Queensland.