Ion-pair assisted recovery of matrix-assisted laser desorption/ionization mass spectral signals from SDS-containing peptide-protein mixtures.
We have developed a simple and effective means of using alkylammonium ion-pairing agents, such as cetyltetramethylammonium bromide, to recover matrix assisted laser desorption/ionization mass spectrometric (MALDI-MS) signals from sodium dodecyl sulfate (SDS)-containing protein and peptide samples. A two-layer method of matrix preparation, with a bottom matrix layer of ion-pairing agents and a top matrix layer of SDS protein samples, is essential for reproducible MALDI mass spectra with good recovery. Both buffer ions and ion-pairing agents have profound effects on signal recovery and can be rapidly and systematically optimized. This practical technique, termed ion-pair assisted recovery (IPAR), is compatible with major SDS-based biotechniques and can be easily incorporated into high-throughput proteomic analysis.
Rajnarayanan
Rajendram V
RV
Laboratory of Muscle Biology, Muscle Proteomics and Nanotechnology Section, National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institute of Health, 50 South Drive, Bethesda, Maryland 20892-8024, USA.
Wang
Kuan
K
D002021:Buffers; D007477:Ions / Q000737:chemistry; D010455:Peptides / Q000032:analysis* / Q000737:chemistry; D011506:Proteins / Q000032:analysis* / Q000737:chemistry; D015203:Reproducibility of Results; D012967:Sodium Dodecyl Sulfate; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D002021:Buffers; D007477:Ions; D010455:Peptides; D011506:Proteins; D012967:Sodium Dodecyl Sulfate