Thermo Fisher LTQ-Orbitraps
The Thermo Fisher LTQ-Orbitrap Elite combines the newest high speed dual linear ion trap and S-lens technology in the LTQ Velos, with the Orbitrap mass analyzer, providing high mass accuracy and resolution for both MS and MSn spectra. This instrument is typically run with the Orbitrap acquiring full MS data at low ppm mass accuracy and resolution up to 120,000 HWFM, while the LTQ acquires collisionally induced dissociation (CID) MSMS scans of peptides for identification and modification site determination. Other operational configurations include SRM/MRM experiments, and high energy collisions (HCD) with high accuracy MSMS spectra in the Orbitrap. Two Elite instruments are currently running in our facility at this time, both configured with capillary chromatographic and nanospray ESI interfaces.
Driving the chromatography on both of the Orbitraps is the Waters nanoACQUITY UPLC system. This system maintains accurate gradient production in a splitless configuration in the sub-μl range at pressures up to 10,000psi. The autosampler allows automated trapping for sample concentration and desalting and provides excellent reproducibility and linearity across the injection volume range of 0.1 to 20μl.
New Objective Inc. provides the nanocapillary chromatography hardware and ESI source used exclusively on the LTQ-Orbitrap. The PicoView source allows easy positioning and visualization of the ESI spray, which is key to operation of the high sensitivity nanocapillary columns with integrated spray tips (Picofrit columns) and trap columns (IntegraFrit columns) provided by this long-time collaborator with our lab. No other chromatographic configuration has demonstrated the versatility, robustness and extremely high sensitivity that these products provide. Also shown is a smart enhancement for nannspray source which was developed here in this lab, the Active Background Ion Reduction Device, or ABIRD, by ESI Source Solutions. THis device isolates the instrument from the highly variable background ions of a busy laboratory, allowing ultra-trace level proteomics analysis of highly complex mixtures of peptides.
Thermo Fisher Triple Quadropole Mass Spectrometer
Thermo Scientific* TSQ Quantum Ultra triple stage quadrupole mass spectrometer is the value-conscious choice for a wide variety of LC/MS applications, including pharmaceutical, environmental, food safety, clinical research, and forensic toxicology. This instrument is available many targeted projects, call to discuss your project first.
Intact Molecular Weight Determinations, proteins, peptides, oligos.
The Bruker MALDI-TOF is a reflectron based instrument used for peptide, DNA/oligo, protein and many other small and large molecular weight determinations. Due to the "soft" ionization of the MALDI source and low charge state typically observed, the mass range can vary from below 100Da to well over 250,000 Da.
Applied Biosystems Procise 494-HT
Edman sequencing is a classic protein characterization technique that has been a mainstay in this laboratory for over 20 years. This process requires >1pmol of protein, either in solution of on PVDF blot, and can quantitatively determine the sequence of one or several protein or peptides with a cycle time (one amino acid per cycle) of about 45 minutes. This is not a database dependent technique, so it can be considered the original form of denovo protein sequencing. Typical sequences are 10 amino acids, but extended sequence runs can be preformed for novel proteins or peptides.