Ohtsu Iwao, Nakanisi Tsuyoshi, Furuta Masaru, Ando Eiji, Nishimura Osamu
Life Science Laboratory, Analytical & Measuring Instruments Division, Shimadzu Corporation, 1 Nishinokyo-Kuwabaracho, Nakagyo-ku Kyoto 604-8511, Japan.
J Proteome Res. 2005 Jul-Aug;4(4):1391-6. doi: 10.1021/pr050073n.
We have developed new procedures to identify proteins after they are detected by Western blotting or other interactions such as lectin blotting on membranes. Our method is based on the combination of on-membrane MALDI-TOF mass spectrometry with piezoelectric chemical inkjet technology. Using this method the GroEL, FtsZ, DnaK, and GroES proteins were successfully identified from Escherichia coli after separation on two-dimensional gels, immunostaining, and on-membrane digestion. A glycoprotein detected by lectin blotting with concanavalin A was also identified using this technique.
我们开发了新的方法，用于在蛋白质通过蛋白质免疫印迹法或其他相互作用（如膜上凝集素印迹法）检测后对其进行鉴定。我们的方法基于膜上基质辅助激光解吸电离飞行时间质谱（MALDI-TOF）与压电化学喷墨技术的结合。使用该方法，在二维凝胶上分离、免疫染色和膜上消化后，成功从大肠杆菌中鉴定出了GroEL、FtsZ、DnaK和GroES蛋白。使用该技术还鉴定了通过伴刀豆球蛋白A凝集素印迹法检测到的一种糖蛋白。
