Mass spectrometry
The IgE-reactive bands of raw fish extracts were excised from the
SimplyBlue SafeStain-stained SDS-PAGE gel, destained and digested with
trypsin following our laboratory’s established
protocol21. The tryptic digests were analyzed on the
UltrafleXtreme MALDI-ToF/ToF System (Bruker) for obtaining the peptide
mass fingerprints. Searches against the entire protein sequence database
(NCBI and UniProt) using the MASCOT search engine were then performed
for protein identification.