title: Optimization of neuropeptide extraction from the mouse hypothalamus.
abstract: Sample preparation for neuropeptidomic studies is a critical issue since protein degradation can produce high levels of peptides that obscure the endogenous neuropeptides. We compared different extraction conditions for the recovery of neuropeptides and the formation of protein breakdown fragments from mouse hypothalami. Sonication and heating in water (70 degrees C for 20 min) followed by cold acid and centrifugation enabled the efficient extraction of many neuropeptides without the formation of protein degradation fragments seen with hot acid extractions. The hot water/cold acid extraction procedure resulted in the reproducible recovery of many hypothalamic peptides, including several novel peptides.
authors.affiliation: Department of Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, New York 10461, USA.
authors.forename: Fa-Yun
authors.initials: FY
authors.lastname: Che
authors.forename: Xin
authors.initials: X
authors.lastname: Zhang
authors.forename: Iryna
authors.initials: I
authors.lastname: Berezniuk
authors.forename: Myrasol
authors.initials: M
authors.lastname: Callaway
authors.forename: Jihyeon
authors.initials: J
authors.lastname: Lim
authors.forename: Lloyd D
authors.initials: LD
authors.lastname: Fricker
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002853:Chromatography, Liquid; D005260:Female; D007031:Hypothalamus / Q000737:chemistry* / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009479:Neuropeptides / Q000302:isolation & purification* / Q000378:metabolism; D053719:Tandem Mass Spectrometry
chemical_list: D009479:Neuropeptides