High-throughput protein digestion by trypsin-immobilized monolithic silica with pipette-tip formula.
Based on the monolithic silica gel materials with hierarchical pore structure and on the SPE devices (MonoTip) developed thereof, a trypsin-immobilized monolithic silica in a pipette tip (MonoTip Trypsin) suitable for digesting proteins has been newly developed. The surface of monolithic silica fixed into the tip was chemically modified with trypsin via an aminopropyl group. Trypsin-immobilized monolith successfully performed a rapid digestion of reduced and alkylated proteins with only a few times pipetting operation for the pre-treatment procedure of chromatographic analysis. The novel solid-phase digestion tool using monolithic silica allows a high-throughput trypsin proteolysis of bio-substances in proteomics.
Ota
Shigenori
S
GL Sciences Inc., 237-2 Sayamagahara, Iruma, Saitama, 358-0032, Japan. sig-ota@gls.co.jp
Miyazaki
Shota
S
Matsuoka
Hideo
H
Morisato
Kei
K
Shintani
Yukihiro
Y
Nakanishi
Kazuki
K
D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D004800:Enzymes, Immobilized; D013058:Mass Spectrometry / Q000379:methods; D010446:Peptide Fragments / Q000302:isolation & purification; D010449:Peptide Mapping / Q000379:methods; D011506:Proteins / Q000302:isolation & purification*; D012710:Serum Albumin, Bovine / Q000302:isolation & purification; D012822:Silicon Dioxide; D014357:Trypsin
D004800:Enzymes, Immobilized; D010446:Peptide Fragments; D011506:Proteins; D012710:Serum Albumin, Bovine; D012822:Silicon Dioxide; D014357:Trypsin