Gharahdaghi F, Kirchner M, Fernandez J, Mische S M
Rockefeller University Protein/DNA Technology Center, New York, New York 10021, USA.
Anal Biochem. 1996 Jan 1;233(1):94-9. doi: 10.1006/abio.1996.0012.
Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), in conjunction with enzymatic digestion of proteins and molecular weight search of peptide-mass database is a powerful technique for peptide/protein identification. Ideally, peptide mixtures should be compatible with both MALDI-TOF and microsequencing. In our laboratory, enzymatic digestion and extraction of peptides from polyvinylidene difluoride (PVDF)-bound proteins is performed in the presence of nonionic detergents. However, nonionic detergents have been shown to cause signal suppression in MALDI-TOF analysis. This study demonstrates that by using a modified matrix solution, peptide-mass fingerprinting of PVDF-bound proteins by MALDI-TOF can be obtained in the presence of nonionic detergents such as hydrogenated Triton X-100 (RTX-100), octylglucopyranoside, and Tween 20.
基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF MS），结合蛋白质的酶解消化和肽质量数据库的分子量搜索，是一种用于肽/蛋白质鉴定的强大技术。理想情况下，肽混合物应与MALDI-TOF和微量测序都兼容。在我们实验室中，从聚偏二氟乙烯（PVDF）结合的蛋白质中酶解消化和提取肽是在非离子去污剂存在的情况下进行的。然而，已表明非离子去污剂会在MALDI-TOF分析中导致信号抑制。本研究表明，通过使用改良的基质溶液，在氢化Triton X-100（RTX-100）、辛基吡喃葡萄糖苷和吐温20等非离子去污剂存在的情况下，可通过MALDI-TOF获得PVDF结合蛋白质的肽质量指纹图谱。
