Difference between revisions of "LX Reference Carvalho 2010"
(→MFQL queries) |
|||
Line 3: | Line 3: | ||
====Materials and Methods==== | ====Materials and Methods==== | ||
− | + | Top-down analysis of the fly total membrane lipidome was performed on an LTQ Orbitrap mass spectrometer (Thermo Fisher Scientific, Bremen, Germany) equipped with a NanoMate robotic nanoflow ion source (Advion BioSciences, Ithaca, NJ, USA) as described previously (Schwudke et al., 2007a). MS/MS experiments on the LTQ Orbitrap and the QSTAR Pulsar i (MDS Sciex, Toronto, Canada) mass spectrometers were performed as described previously (Schwudke et al., 2007b). Absolute quantification of ceramides and hexosyl-ceramides was performed by adding 250 nmol of Cer 35:1;2 and GlcCer 30:1;2 (Avanti Polar Lipids. Alabaster, AL, USA) as standards to the lipid extracts. Quantification of sphingolipids relied on extracted intensities of specific structural fragments of the long chain base (LCB): m/z 264.2 (for LCB 18), 236.2 (for LCB 16), 234.2 (for LCB 16:1), 208.2 (for LCB 14) and 206.2 (for LCB 14:1). | |
====MFQL queries==== | ====MFQL queries==== | ||
− | MFQL queries for ceramides, PE-ceramids and glucosylceramide ( | + | MFQL queries for ceramides, PE-ceramids and glucosylceramide (hexosyl ceramides) in positive mode: |
* [[Cer_MRM.mfql | Cer_MRM.mfql]] | * [[Cer_MRM.mfql | Cer_MRM.mfql]] | ||
* [[CerPE_MRM.mfql | CerPE_MRM.mfql]] | * [[CerPE_MRM.mfql | CerPE_MRM.mfql]] | ||
* [[HexCer_MRM.mfql | HexCer_MRM.mfql]] | * [[HexCer_MRM.mfql | HexCer_MRM.mfql]] |
Revision as of 16:59, 24 February 2012
Queries, spectra and results
Materials and Methods
Top-down analysis of the fly total membrane lipidome was performed on an LTQ Orbitrap mass spectrometer (Thermo Fisher Scientific, Bremen, Germany) equipped with a NanoMate robotic nanoflow ion source (Advion BioSciences, Ithaca, NJ, USA) as described previously (Schwudke et al., 2007a). MS/MS experiments on the LTQ Orbitrap and the QSTAR Pulsar i (MDS Sciex, Toronto, Canada) mass spectrometers were performed as described previously (Schwudke et al., 2007b). Absolute quantification of ceramides and hexosyl-ceramides was performed by adding 250 nmol of Cer 35:1;2 and GlcCer 30:1;2 (Avanti Polar Lipids. Alabaster, AL, USA) as standards to the lipid extracts. Quantification of sphingolipids relied on extracted intensities of specific structural fragments of the long chain base (LCB): m/z 264.2 (for LCB 18), 236.2 (for LCB 16), 234.2 (for LCB 16:1), 208.2 (for LCB 14) and 206.2 (for LCB 14:1).
MFQL queries
MFQL queries for ceramides, PE-ceramids and glucosylceramide (hexosyl ceramides) in positive mode: