Identification of tubulin peptides by ProFound peptide mass fingerprinting using immunoprecipitation experiments and averaged MALDI-TOF/TOF data.

Tubulin was immunoprecipitated from HEK293 cell lysate using a rabbit polyclonal antibody and Protein A/G Dynabeads. After isolation, all eluted proteins were reduced and alkylated, digested with trypsin, and analyzed on an ABI 4800 MALDI-TOF/TOF mass spectrometer. Multiple individual spectra were acquired with internal calibration and between 10 and 23 individual measurements for each peptide were used for calculating the average observed masses for each peptide. Mass Tolerance is in parts-per-million (ppm), Peptide Set defines the peptides included for search and Search Mass Range/pI Range are input parameters for Profound. Top Protein ID and Expectation Value were calculated within ProFound from the mass spectrometry data using the IPI Human database (2010-02-01).
Identification of tubulin peptides by ProFound peptide mass fingerprinting using immunoprecipitation experiments and averaged MALDI-TOF/TOF data.