The removal of Triton X-100 by dialysis is feasible!

被引:7
作者
Opitz, Stefan [1 ]
Hannika, Franziska [1 ]
Krueger, Thomas [2 ]
Rhode, Heidrun [1 ]
机构
[1] Jena Univ Hosp, Inst Biochem 1, D-07743 Jena, Germany
[2] Hans Knoll Inst Jena, Leibniz Inst Nat Prod Res & Infect Biol, Dept Mol & Appl Microbiol, D-07745 Jena, Germany
关键词
Hoechst; 33342; Critical micellar concentration; Sample preparation; Microdialysis; Particle size distribution; MALDI-TOF; CRITICAL MICELLE CONCENTRATION; MASS-SPECTROMETRY; ALKALINE-PHOSPHATASE; NONIONIC SURFACTANTS; SAMPLE PREPARATION; ORGANIC ADDITIVES; SPIN COLUMN; CLOUD POINT; DETERGENTS; PROTEIN;
D O I
10.1007/s00216-014-8333-3
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Triton X-100 has been widely used in many analytical and preparative protocols for a long time. Nevertheless, mass spectrometry, chromatographic separation, and spectrophotometric readout may be considerably hampered by this detergent due to signal suppression, complex formation, and high blank values, respectively. Additionally, Triton X-100 is not safe to remove prior to analytics. Here, microdialysis is introduced as a parallelizable, high-throughput method to clean samples from Triton X-100 with high efficacy and precision. To achieve this, we exploit the potential to considerably increase the critical micellar concentration of Triton X-100 by alteration of matrix properties. To that end, addition of several chaotropic compounds and organic solvents has been shown to increase the critical micellar concentration as well as the removal rate of the detergent. For application, matrix additives can be selected for analyte stability requirements out of a variety of compounds. Conveniently, all these additives are removable subsequently using the same microdialysis tool for downstream analytics requirements. Applicability and protocols are shown with proteomic sample preparation of purified proteins and complex protein mixtures prior to matrix-assisted laser desorption ionization (MALDI) mass spectrometry.
引用
收藏
页码:1107 / 1118
页数:12
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