The samples were added to 5 µl of 0.1% TFA for re-suspension. The TFA was mixed at a 1:1 ratio with α-cyano-4-hydroxy-trans-cinnamic acid in 50% ACN. The mixed solution (1 µl) was added to a sample target plate. Using an ABI 4800 MALDI TOF/TOF Plus mass spectrometer, peptide MS and MS/MS were performed (Applied Biosystems; Thermo Fisher Scientific, Inc., Waltham, MA, USA). Using GPS Explorer™ software (version 3.6; Applied Biosystems; Thermo Fisher Scientific, Inc.), a search (MS+MS/MS) was performed. The time of flight spectra recorded the positive ion reflector mode in a mass range of 800-4,000 Da. According to the MS and MS/MS spectra, proteins were successfully determined, with a confidence interval of ≥95%, using the Mascot V2.3 search engine (Matrix Science, London, UK). The other parameters were set as follows: NCBI-Animals database; 100 ppm for precursor ion tolerance; fixed modifications of carbamidomethyl; 0.3 Da for fragment ion tolerance; and partial modifications of acetyl and oxidation.
Abi 4800 maldi tof tof plus mass spectrometer
Manufactured by Thermo Fisher Scientific
Sourced in United States
The ABI 4800 MALDI TOF/TOF Plus mass spectrometer is a flexible and versatile instrument designed for high-throughput proteomics and protein analysis. It utilizes matrix-assisted laser desorption/ionization (MALDI) and tandem time-of-flight (TOF/TOF) technology to provide accurate mass measurements and detailed structural information on biomolecules.
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2 protocols using abi 4800 maldi tof tof plus mass spectrometer
1
MALDI-TOF/TOF for Protein Identification
2
MALDI-TOF/TOF Protein Identification Protocol
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In gel digestion was performed according to Katayama et al. [21] . After being completely dried, the D-shape larval mussel samples were re-suspended with 5 mL of 0.1% TFA followed by mixing in 1:1 ratio with a saturated solution of a-cyano-4-hydroxy-trans-cinnamic acid in 50% acetonitrile [22] . One microliter of mixture was analyzed by an ABI 4800 MALDI-TOF/TOF Plus mass spectrometer (Applied Biosystems, Foster City, USA), data were acquired in a positive MS reflector using a CalMix5 standard to calibrate the instrument (ABI4800 Calibration Mixture). Both the MS and MS/MS data were integrated and processed using the GPS Explorer V3.6 software (Applied Biosystems, USA) with default parameters. Proteins were successfully identified based on 95% or higher confidence interval of their scores in the MASCOT V2.4 search engine (Matrix Science Ltd., London, U.K.). The following parameters were used in the search: NCBInr Metazoa (Animals) (2861494 sequences) database; trypsin as the digestion enzyme; one missed cleavage site; partial modifications of cysteine carbamidomethylation and methionine oxidization; no fixed modifications; 0.15 Da for precursor ion tolerance and 0.25 Da for fragment ion tolerance. Individual ions scores >40 indicate identity or extensive homology (P < 0.05).
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