Our protein ID procedure is the following:
- In-gel Digestion
1D gel bands or 2D gel spots are processed for in-gel trypsin digestion. The resulted tryptic peptides are extracted, dried down, and re-suspended for LC-MS/MS analysis.
- LC-MS/MS Analysis
Tryptic peptides from the above step are run on a nano HPLC with a 75 μM ID reverse phase column, which is connected to a Thermo LTQ ion trap mass spectrometer. Data from a run of 45 minutes will be collected and used for protein database search.
- Protein Database Search
LC-MS/MS data are searched against protein databases such as NCBI or SwissProt. A protein hit list will be generated.
- Manual Validation
The initial hit list will be manually examined by our scientists to eliminate any false positives based on LC-MS/MS raw data, bioinformatics analysis, and sample information. We only provide protein ID with high confidence level.
- Report
A Protein ID Report with a list of all identified proteins is delivered to customer by email.
