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  • Identification and relative quantification of neuropeptides from the endocrine tissues.

Identification and relative quantification of neuropeptides from the endocrine tissues.

Methods in molecular biology (Clifton, N.J.) (2009-12-17)
Kurt Boonen, Steven J Husson, Bart Landuyt, Geert Baggerman, Eisuke Hayakawa, Walter H M L Luyten, Liliane Schoofs
ABSTRACT

Endocrine tissues like the pituitary, hypothalamus and islets of Langerhans are rich in bioactive peptides. These are used for intercellular signalling and are involved in regulation of almost all physiological processes. Peptidomics is the comprehensive analysis of peptides in tissues, fluids and cells. Peptidomics applied to (neuro-)endocrine tissues aims therefore to identify as many bioactive peptides as possible. Peptidomics of (neuro-)endocrine tissues requires an integrated approach that consists of careful sample handling, peptide separation techniques, mass spectrometry and bioinformatics. Here we describe the methods for isolation and dissection of endocrine tissues, the extraction of bioactive peptides and further sample handling and identification of peptides by mass spectrometry and hyphenated techniques. We also present a straightforward method for the comparison of relative levels of bioactive peptides in these endocrine tissues under varying physiological conditions. The latter helps to elucidate functions of the bioactive peptides.

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Protease Inhibitor Cocktail powder, for use with bacterial cell extracts, lyophilized powder