The protein sample (20 μl at 10 pM/μl) was desalted on-line through a Waters MassPREP column (2.1 mm × 10 mm), eluted with a gradient from solvent A [2% CH3CN, 97% H2O, 1% formic acid (FA)] to solvent B (97% CH3CN, 2% H2O, 1% FA), and delivered to a Waters LCT electrospray ionization mass spectrometer precalibrated using myoglobin. An envelope of multiple charged signals was obtained in positive ionization from 500 to 2000 mass/charge ratio (m/z) and deconvoluted using MaxEnt 1 software over the range of 44–48 kDa to 1 Da resolution to give the molecular mass of the protein.

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