title: Removal of detergents from proteins and peptides in a spin-column format.
abstract: To enable downstream analysis, it is critical to remove unbound detergents from protein and peptide samples. This unit describes the use of a high-performance resin that offers exceptional detergent removal for proteins and peptides. The easy-to-use spin format significantly improves results over the standard drip column and batch methodologies, with >95% removal of 1% to 5% detergents, including SDS, sodium deoxycholate, CHAPS, Triton X-100, Triton X-114, NP-40, Brij-35, octyl glucoside, octyl thioglucoside, and lauryl maltoside, with high recovery of proteins and peptides. Detergent removal efficiency is evaluated using colorimetric methods and mass spectrometry (MS). BSA tryptic peptides have been successfully analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and matrix-assisted laser desorption/ionization (MALDI)-MS for identification of protein, following detergent removal using the resin. Advantages of this method include speed (less than 15 min), efficient detergent removal, and high recovery of proteins and peptides.
authors.affiliation: Thermo Fisher Scientific, Research & Development, Rockford, Illinois.
authors.forename: Babu S
authors.initials: BS
authors.lastname: Antharavally
mesh_terms: D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity / Q000191:economics / Q000379:methods*; D002853:Chromatography, Liquid; D003902:Detergents / Q000302:isolation & purification*; D006801:Humans; D010455:Peptides / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry*; D012710:Serum Albumin, Bovine / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053719:Tandem Mass Spectrometry
chemical_list: D003902:Detergents; D010455:Peptides; D011506:Proteins; D012710:Serum Albumin, Bovine