1. Gel treatment: Gel is washed multiple
times to remove staining dye and other chemicals inhibitory
to Mass Spectrometry
2. In-gel trypsin digestion: Trypsin
digestion buffer is added and protein is digested in gel
at 37°C
3. Peptide extraction: Digested peptide
sample is extracted from gel by extraction buffer
4. Desalting: The digested tryptic
peptides are desalted by C-18 Zip-tip (Millipore)
5. Spotting: Sample is mixed with matrix
buffer and spotted on MALDI plate
6. MALDI-TOF: MS spectra are obtained
using ABI4700/ABI4800 proteomic analyzer
7. MALDI-TOF/TOF: Ten to twenty most
abundant peptides are further subjected to fragmentation
(MS/MS)
8. Database search: Both MS and MS/MS
spectra are submitted for database search using GPS explorer
equipped with MASCOT search engine
9. Data Report (see a sample
report of protein ID analysis)
Call our tech support at 510-887-0889 or send email
to support@appliedbiomics.com
with any technical questions.
