Proteomics meets genetics: SILAC labeling of drosophila melanogaster larvae and cells for in vivo functional studies
Capitolo
Data di Pubblicazione:
2014
Citazione:
Proteomics meets genetics: SILAC labeling of drosophila melanogaster larvae and cells for in vivo functional studies / A. Cuomo, R. Sanfilippo, T. Vaccari, T. Bonaldi (METHODS IN MOLECULAR BIOLOGY). - In: Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) : Methods and Protocols / [a cura di] B. Warscheid. - [s.l] : Humana Press Inc., 2014. - ISBN 9781493911417. - pp. 293-311 [10.1007/978-1-4939-1142-4_21]
Abstract:
Stable isotope labeling by amino acids in cell culture (SILAC) is an established and potent method for quantitative proteomics. When combined with high-resolution mass spectrometry (MS) and efficient algorithms for the analysis of quantitative MS data, SILAC has proven to be the strategy of choice for the in-depth characterization of functional states at the protein level. The fruit fly Drosophila melanogaster is one of the most widely used model systems for studies of genetics and developmental biology. Despite this, a global proteomic approach in Drosophila is rarely considered. Here, we describe an adaptation of SILAC for functional investigation of fruit flies by proteomics: We illustrate how to perform efficient SILAC labeling of cells in culture and whole fly larvae. The combination of SILAC, a highly accurate global protein quantification method, and of the fruit fly, the prime genetics and developmental model, represents a unique opportunity for quantitative proteomic studies in vivo.
Tipologia IRIS:
03 - Contributo in volume
Keywords:
Quantitative proteomics; Drosophila melanogaster; SILAC; Schneider cell; Saccharomyces cerevisiae; Intact organism labeling; Mass spectrometry
Elenco autori:
A. Cuomo, R. Sanfilippo, T. Vaccari, T. Bonaldi
Link alla scheda completa:
Titolo del libro:
Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) : Methods and Protocols