Optimization of neuropeptide extraction from the mouse hypothalamus.
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.
Che
Fa-Yun
FY
Department of Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, New York 10461, USA.
Zhang
Xin
X
Berezniuk
Iryna
I
Callaway
Myrasol
M
Lim
Jihyeon
J
Fricker
Lloyd D
LD
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
D009479:Neuropeptides