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[分子量測定]Molecular weight measurement本服務主要對核酸、蛋白質、胜肽作精確分子量測定，樣品濃度至少需要5pmol/ul以上。 [蛋白質身份鑑定]Protein Identification by LC-MS/MS本服務以LC-ESI/MS/MS對蛋白質消化產物胜肽進行胜肽質量與胜肽碎裂碎片質量量測，並與資料庫比對而得蛋白質身份。此方法適合50個蛋白質以下之混合物，例如一維膠體電泳帶，樣品量建議10ng以上。 Large-scale Profiling of Protein Identification by 2D LC-MS/MS本服務以2D-LC-ESI/MS/MS對蛋白質消化產物胜肽進行胜肽質量與胜肽碎裂碎片質量量測，並與資料庫比對而得蛋白質身份。此方法適合50個蛋白質以上之蛋白質混合物，例如: total lysate, sub-proteome等。胜肽混合物先以第一維強陽離子交換層析法粗分離並分液收集約25～40管，接著將各管進行Zip-tip去鹽，進行第二維C18液相層析電灑串連質譜法進行胜肽質量與胜肽碎裂碎片質量量測，並與資料庫比對而得蛋白質身份。收費標準以分液收集個數(collected fractions)乘以重複分析次數計算。因總儀器分析時間冗長，欲使用此項服務者，請預先與核心設施預約儀器時間。 [蛋白質轉譯後修飾鑑定]Identification of post-translational Modifications本服務以LC-ESI/MS/MS對轉譯後修飾蛋白質消化產物胜肽進行胜肽質量與胜肽碎裂碎片質量量測，並與資料庫比對而得蛋白質身份及修飾點位置資訊。此方法適合純化後蛋白質，例如50個蛋白質以下之混合物。欲取得較多修飾點位置資訊必須有較高的蛋白質序列涵蓋率，因此蛋白質越純、量越多，效果越好！依修飾程度差異，至少需要2~10ug以上的蛋白質。磷酸化部分，本核心設施提供TiO2磷酸化胜肽純化法之相關耗材、試劑及操作手冊供使用者自行操作，亦對初學者提供教育訓練教學課程，請多加利用！欲使用此服務者，請預先與核心設施聯絡，詳談樣品處理步驟及預約儀器時間。 Large-scale Phosphoproteome with IMAC enrichment本服務以LC-ESI/MS/MS對磷酸化蛋白質消化產物胜肽進行胜肽質量與胜肽碎裂碎片質量量測，並與資料庫比對而得蛋白質身份及磷酸化修飾點位置資訊。此方法適合50個蛋白質以上之蛋白質混合物，例如: total lysate, sub-proteome等。由於胜肽複雜度相當高，必須結合IMAC進行磷酸化胜肽純化，才能有效提取磷酸化資訊。欲取得較多修飾點位置資訊，起始蛋白量越多，效果越好！依修飾程度差異，至少需要數百ug～數mg以上的蛋白質。此部分，本核心設施提供代做服務，欲使用此服務者，請預先與核心設施聯絡，詳談樣品處理步驟及預約儀器時間。 [訓練課程]以下課程內容提供線上教學影帶，並於每年三月及九月舉辦教育訓練課程，報名資訊請參閱教育訓練分頁。 |
This service is mainly to measure the accurate molecular weight of nucleic acids, proteins, and peptides by MALDI-TOF. The concentration of samples should be more than 5pmol/ul.
This service is to measure the peptide mass and peptide fragment mass for the peptide of protein digestion products by LC-ESI/MS/MS and to identify the proteins with matching to database. The mixed samples composed of less than 50 proteins could use this method, such as 1D gel electrophoresis, and recommended the amount of sample should be more than 10ng.
This service is to measure the peptide mass and peptide fragment mass for the peptide of protein digestion products by 2D-LC-ESI/MS/MS and to identify the proteins with matching to database. The mixed samples composed of less than 50 proteins could use this method, such as total lysate, sub-proteome etc. First, the peptide mixture should be separated roughly by strong cation exchange chromatography and collect liquid fraction of 25~40 tubes. After that, each liquid of tube must be desalted with ZipTip to measure the peptide mass and peptide fragment mass by reverse phase C18 liquid chromatography-electrospray tandem mass spectrometry, furthermore to identify the protein with matching to database. The cost is calculated with the number of collected fractions multiplied by the number of repeat analysis. Please make a reservation for the equipment to the core facility early if you would like to use this service, because the whole analyses take a lot of time.
This service is to measure the peptide mass and peptide fragment mass for the post-translational modified protein digestion products by LC-ESI/MS/MS, then to identify the proteins and get the information of modification site with matching to database. The purified proteins could use this method, such as the mixtures composed of less than 50 proteins. To obtain more information of modification site, the protein must have higher sequence coverage. Therefore, the purer and more protein, the more effect! According to the variation of modification, the protein must be needed at least 2~10ug. About the phosphorylation, the core facility supplied the related consumables, reagents and protocol of TiO2 phosphopeptide enrichment for users. In addition, the core facility provided the training courses for the beginners, please feel free to utilize! Please contact with the core facility in advance for the processing of sample treatment and making reservation for the equipment if you would like to use this service.
This service is to measure the peptide mass and peptide fragment mass for the phosphorylated protein digestion products by LC-ESI/MS/MS, then to identify the proteins and get the information of phosphorylated modification site with matching to database. The mixed samples composed of more than 50 proteins could use this method, such as total lysate, sub-proteome etc. It must be combined with IMAC to process phosphopeptide enrichment due to the peptides are very complex. To obtain more information of modification site, the more protein, the more effect! According to the difference of modification level, at least hundreds ug to several mg of proteins were needed. The core facility provide the operation service of IMAC enrichment, so please contact with the core facility in advance for the processing of sample treatment and making reservation for the equipment if you would like to use this service.
The following courses have been made an online instructional video, and the core will hold training courses in March and September each year. About the registration information, please see the training page.