Step |
Name |
Description |
1
|
Marelli_protocol (LCMS2)
|
Prepare and run LCMS, producing one mzXml file per input sample.
|
10
|
Marelli_protocol Sample Prep
|
An organellar 20KgP fraction was subjected to isopycnic density gradient centrifugation and analyzed
by SDS-PAGE and Coomassie blue staining. Fractions enriched for peroxisomes (EP) or mitochondria (M) were identified by Western blotting. Equal amounts of protein derived from each of the hypotonically lysed M and EP fractions were combined and analyzed by ICAT MS/MS.
|
30
|
Marelli_protocol LC MS2
|
ICAT-labeled peptides were analyzed by µLC-ESI-MS/MS. Peptides were separated by on-line reversed-phase chromatography using a 75 µm x 10 cm self-packed Magic C18AQ column (Michrom BioResources) at a flow rate of 300 nL/min. Peptide fragmentation by collision-induced dissociation was performed in an automated fashion using the dynamic-exclusion option from the full-range MS spectra or by GPFm/z using an ion trap mass spectrometer (Thermofinnigan; Yi et al., 2002).
|
40
|
Convert to mzXML
|
|
1000
|
Mark Run Outputs
|
|