Prefractionation methods for individual adult fruit fly hemolymph proteomic analysis

Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Qi ZengScott A Shippy

Abstract

The analysis of blood provides in depth chemical information of physiological states of organisms. Hemolymph (blood) is the fluid in the open circulatory system of Drosophila melanogaster that is the medium for molecules regulating a wide variety of physiological activities and signaling between tissues. Adult Drosophila is typically less than 3mm in length and, as a consequence, the available volume of hemolymph is usually less than 50 nL from individual flies. Proteomic analysis of volume-limited hemolymph is a great challenge for both sample handling and subsequent mass spectrometry characterization of this chemically diverse biological fluid with a wide dynamic range of proteins in concentrations. Less abundant proteins, in particular, could be easily lost during sample preparation or missed by current mass spectrometry methods. This article describes simple and customized RPLC column and IEX columns to prefractionate volume-limited hemolymph without excessive dilution. Step-gradient elution methods were developed and optimized to enhance the identification of novel proteins from an individual fruit fly hemolymph sample. Fractions from each step gradient was analyzed by an Agilent nano-RPLC chip column and then characterize...Continue Reading

References

Mar 25, 2000·Science·M D AdamsJ C Venter
Mar 24, 2000·Science·G M RubinD A Harvey
Oct 19, 2001·Proceedings of the National Academy of Sciences of the United States of America·E De GregorioB Lemaitre
Mar 1, 2003·Nature Reviews. Genetics·Monica Driscoll, Beate Gerstbrein
May 3, 2003·Biochemical and Biophysical Research Communications·Evy VierstraeteLiliane Schoofs
Oct 7, 2004·The Journal of Biological Chemistry·Christine KarlssonUlrich Theopold
Jan 20, 2005·Electrophoresis·Pier Giorgio RighettiEgisto Boschetti
Jan 16, 2008·Analytical Chemistry·Sujeewa C PiyankarageScott A Shippy
Oct 25, 2008·Nucleic Acids Research·Susan TweedieUNKNOWN FlyBase Consortium
Apr 4, 2009·Proteomics·Nobuaki Takemori, Masa-Toshi Yamamoto
Nov 18, 2009·Chemical Senses·Robert R H Anholt, Taufika Islam Williams
Apr 27, 2012·Analytical Chemistry·Sujeewa C PiyankarageScott A Shippy
Nov 20, 2012·Nucleic Acids Research·UNKNOWN UniProt Consortium
Jan 18, 2015·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Qi ZengScott A Shippy

❮ Previous
Next ❯

Citations

Mar 10, 2018·Analytical Chemistry·Marissa R CabayScott A Shippy

❮ Previous
Next ❯

Related Concepts

Related Feeds

ATP Synthases

ATP synthases are enzymes located in the inner mitochondrial membrane that catalyze the synthesis of ATP during cellular respiration. Discover the latest research on ATP synthases here.