| 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 Name | Dr. John Yates, III |
| Title | Professor |
| E-mail | jyates@scripps.edu |
| Address | The Scripps Research Institute
10550 N Torrey Pines Rd.
La Jolla, CA 92037 |
| Country | USA |
| Lab URL | http://fields.scripps.edu/ |
| Data submitted by | Dr. Greg Cantin & Dr. Tao Xu |
| Title | Postdoctoral Fellow |
| E-mail | taoxu@scripps.edu |
| Published/Unpublished |
Published
|
| Journal name |
|
| PubMed ID |
16396503
|
| Sample source |
|
| 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 |
|
| 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 |
|
