Maldi ms application launchpad 2

Total of 143 individual harpacticoids (138 Tigriopus and 5 Schizopera) were transferred to a 6-well plate a day before and fasted. On the day of the experiment, copepods were fixed in 99% ethyl alcohol for approximately 30 min, and then transferred to a 1.5 ml tube containing small amount of ethyl alcohol (0.5 µl). Once all ethyl alcohol in the tube evaporated, leaving the copepod in completely dried state, 4 µl of α-Cyano-4-hydroxycinnamic acid (HCCA) matrix [Acetonitrile 50%; U.P. Water 47.5%; Trifluoroacetic acid 2.5%; supersaturated HCCA (30 mg for total 1 ml matrix)] was added to each tube and incubated at room temperature for at least 20 min. Two microliter of this solution was placed on the target plate of the MALDI-TOF MS equipment (AXIMA Confidence MALDI TOF-Mass Spectrometer; Shimadzu) and once the solution dried completely to crystalize, the plate was placed in the instrument to be analyzed.
Protein mass spectra were measured between 2k to 20k Dalton on a MALDI-MS Application Launchpad 2.9.2 (Shimadzu Biotech) software using linear mode with laser power 80. To create a sum spectrum, 150 profiles were summed up. Each profile was measured 10 times, and the obtained individual protein spectra were exported to ASCII format files after range editing in Data Explorer (TM) software version 4.3.