title: Stable isotope labeling by amino acids in cultured primary neurons.
abstract: Cultured primary neurons are a well-established model for the study of neuronal function. Conventional stable isotope labeling with amino acids in cell culture (SILAC) requires nearly complete metabolic labeling of proteins and therefore is difficult to apply to cultured primary neurons, which do not divide in culture. Here we describe a protocol that utilizes a multiplex SILAC labeling strategy for primary cultured neurons. In this strategy, two different sets of heavy amino acids are used for labeling cells for the different experimental conditions. This allows for a straightforward SILAC quantitation using partially labeled cells because the two cell populations are always equally labeled.
authors: Department of Biochemistry and Molecular Pharmacology and Kimmel Center for Molecular Medicine at the Skirball Institute, New York University School of Medicine, 540 First Avenue, Floor 5, Room Lab 18, New York, NY, 10016, USA.
authors: Guoan
authors: G
authors: Zhang
authors: Katrin
authors: K
authors: Deinhardt
authors: Thomas A
authors: TA
authors: Neubert
mesh_terms: D000596:Amino Acids / Q000737:chemistry*; D000818:Animals; D001921:Brain / Q000166:cytology; D002478:Cells, Cultured; D004210:Dissection; D005260:Female; D007553:Isotope Labeling / Q000379:methods*; D013058:Mass Spectrometry; D009474:Neurons / Q000166:cytology* / Q000378:metabolism*; D011247:Pregnancy; D040901:Proteomics / Q000379:methods*; D051381:Rats
chemical_list: D000596:Amino Acids