Experimental Details - HuPA_00016
Experiment type Mass spectrometry
Short description TNF phosphoproteome anlaysis
Experimental description HeLa cells were grown in both 14N-media and stimulated with TNF, and grown in 15N-media and left unstimulated. Separately, protein lysates were prepared via Trizol reagent, reduced, alkylated, and digested with trypsin. Digests were then mixed 1:1 (14N:15N sample) and subjected to a two step phosphopeptide enrichment of gel-free IEF followed by IMAC. Enriched phosphopeptides were then subjected to a two hour LC-MS/MS/MS (neutral loss triggered MS3) run on an LTQ ion trap MS.
Principal Investigator's NameDr. John Yates, III
AddressThe Scripps Research Institute
10550 N Torrey Pines Rd.
La Jolla, CA 92037
Data submitted byDr. Greg Cantin & Dr. Tao Xu
TitlePostdoctoral Fellow
Published/Unpublished Published
Journal name
PubMed ID 16396503
Sample source
Cell line: HeLa
Source organism Homo sapiens [Taxonomy:9606]
Labeling technique 15N-labeled amino acids
Protease used Trypsin
Is the sample from in gel No
Reduction and Alkylation TCEP and iodoacetamide
Mass spectrometer
Instrument: LTQ [PSI:1000447]
Vendor: ThermoFinnigan [PSI:1000125]
Ionization method ESI [PSI:1000073]
Fragmentation method CID [PSI:1000133]
Mass tolerance used for database searching (MS) 3 Da
Mass tolerance used for database searching (MS/MS) 0.5 Da
Database used for searching IPI version 3.04
Search engine used SEQUEST
Download MS raw dataset  
Download processed files  

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