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.