Gel Washing &
In-gel Trypsin Digestion



Desalting w/ Zip-tip C18 (Millipore)



Spotting on MALDI plate

MALDI-TOF



MALDI-TOF/TOF

Database Search for Protein ID

 

 

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.