核心設施 Core Facilities

蛋白質體核心設施 Proteomics Core Facility

部門介紹 INTRODUCTION

蛋白質體核心設施主要提供以質譜分析技術為主的蛋白質分析服務。本設施採專人操作，服務包含機器操作、數據分析，並提供相關技術諮詢。服務項目包含蛋白質定性分析及蛋白質體相對表現定量分析。定性分析部分包含蛋白質、胜肽等分子量測定，及蛋白質身份鑑定、轉譯後修飾點鑑定。定量分析部分提供包含多種同位素標記定量法及免標記定量法。此外，也提供多項樣品前處理的實體及線上訓練課程。

 

核心設施位置：N341

電話：02-27899136

E-mail: mass@ibms.sinica.edu.tw

                                                                     

 

Welcome to the IBMS Proteomics Core Facility. The Proteomics Core Facility (PCF) provides a myriad of services for protein qualitation and quantitation using mass spectrometry based proteomics approaches. The PCF is professionally staffed with installed equipment such as state-of-the-art mass spectrometers, nanoUPLC systems, and data processing tools. Our services cover protein identification (for both simple and highly complex mixture), characterization of protein post-translational modifications and large-scale proteome quantitation. The PCF also provide advice for sample preparation, experimental design and preliminary data interpretation.

 

Contact us:

Phone: 02-27899136

E-mail: mass@ibms.sinica.edu.tw

負責人
電話: 02-26523926
職務名稱: 負責人 (Director)
  1. 實驗室管理
  2. 技術諮詢
  3. 分析方法建立、開發

成員 Staff
職務名稱: 實驗室技術人員 (Technician)
  • 樣品分析
  • 毒化物安全管理

職務名稱: 實驗室技術人員 (Technician)
  • 樣品分析

 

 

 

 

  致謝說明
1. 若您使用核心設施後有相關之成果發表(期刊、會議摘要、學位論文等)，請記得於文章致謝中提及設施名稱及服務。
2. 若設施人員的協助對您的成果有重要貢獻，也請您考慮將相關人員列名為共同作者。
3. 文獻紀錄為核心設施爭取設備更新、人員考績和升等的重要依據，感謝您的支持。
4. 請將文獻資訊(期刊、年份、頁數及著作題目)，e-mail告知公用儀器室(E-mail: CEC@ibms.sinica.edu.tw) 供年度文獻統計用。


蛋白質體核心設施：Proteomics Core Facility

例如：…thank the Proteomics Core Facility of the Institute of Biomedical Sciences, 
Academia Sinica for the LC/MS/MS analysis.

 

文獻下載

Acknowledgment
Acknowledgment or authorship for the Core lab's contribution is crucial in supporting the Core lab's ongoing operations and development, as well as the acquisition of future instrumentation and capabilities. In the case of acknowledgment, please send to the Core Facilities lab the bibliographical information of your publications (E-mail: CEC@ibms.sinica.edu.tw). Many thanks.

 

The name of the Core Facility: Proteomics Core Facility

 E.g.：…thank the Proteomics Core Facility of the Institute of Biomedical Sciences, Academia Sinica for the LC/MS/MS analysis.

 

 

Publication

 

1. 是否可委由核心設施代為進行樣品前處理(In-solution/in-gel digestion,etc)操作？
因人力限制，核心設施提供教育訓練及Protocol 下載，不提供代做服務。初學者可至教育訓練分頁觀看線上教學影帶，實體教育訓練於每年三月及九月舉辦，相關資訊刊登於教育訓練分頁，請依說明進行報名。
 
2. 申請前往核心設施進行In-gel digestion 前，樣品應該準備到什麼程度?
請將膠體挖取下來（小心避免任何keratin 的污染源），放置eppondorf 保存再帶至核心設施進行In-gel digestion 實驗。
 
3. 至核心設施進行實驗需要自備試劑？
核心設施提供相關試劑與耗材，請愛惜資源並小心避免試劑污染。
 
4. 以 Sliver stain 方式的樣品，是否可送測分析？
普通 Sliver stain 方法中含有aldehyde，恐會影響質譜分析之performance，故不適宜進行質譜分析，對於低濃度之樣品，本核心設施建議採用具高敏感度且快速的SYPRO Ruby染色方式，此染色方式將不影響質譜分析效果。對於Sliver染色方式之各項分析條件本核心設施正積極規劃測試中。
 
5. 如何估計送樣之蛋白質含量？
請使用適當的蛋白質定量方法估算，並於送樣時告知您使用的定量法。若是預計要進行In-solution digestion 的樣品，本核心提供μBCA 試劑組供蛋白質定量。In-gel digestion的樣品建議可於同一片膠內同時跑一個已知量的內標蛋白質，例如BSA，可供染色後比對估算膠內蛋白質含量。請注意！此估計量會後續胜肽萃取、去鹽等步驟而有不同程度的損失，請小心操作後續程序以取得高回收率。
 
6. 樣品送測LC-MS/MS 前之樣品製備注意事項為何？
請參考送測流程分頁內「送測樣品注意事項」。
 
7. 如何查詢樣品的分析進度？
原則上會依循樣品收件順序進行上機分析，但本核心設施依分析容量、樣品性質及分析所需時間保留部分調控權，若對進度有疑問者，可電洽核心設施。
(如遇儀器故障、維護保養須延遲服務所需時間，將於網頁最新訊息公佈狀態)
 
8. 如何取得樣品的實驗數據？
服務完成時，實驗數據會以E-mail 方式同時寄送至主持人與送件聯絡人之電子信箱中，若檔案過大，會上傳至各實驗室所屬FTP 空間，並以E-mail 及電話通知下載。
 
9. 可取回分析剩餘樣品嗎？
分析剩餘量約為原始量之10％～50％，本設施保留分析剩餘樣品3 個月，若申請者欲索回剩餘樣品，請務必於3 個月內提出要求。
 
10. 核心設施是否會幫送件者保存所有實驗數據？
實驗原始數據從寄出數據給送件者起，僅於核心設施資料庫內保留一年，如需存取原始數據資料者，敬請送件者自行注意到期時限並於期限前通知核心設施進行數據存取。因核心設施之儲存系統有限，所有數據一旦超過保留時限後一律刪除，無法提供數據查詢或下載。
 
11. 如何申請蛋白質表現相對定量分析服務?
本服務一律須優先與核心設施相關人員進行諮詢與討論，經確認儀器預約時間及費用，再行送件申請程序。(註：本服務因為質譜儀需向生化所核心設施預定儀器使用時間，欲使用此技術請至少提前三個月預約)
 
12. 可否指定使用哪一種機型進行分析服務？
本核心設施實驗服務人員會考量儀器條件與狀況，選擇適合的質譜儀提供分析服務，以達到高效率的服務速度，所以無法由使用者指定特定機型進行分析服務，敬請配合與諒解。
 
13. 對送測樣品的結果有疑問，該與誰聯絡？
若對樣品實驗結果有任何疑問，請洽本核心設施，我們將為您做詳盡的解說。
 
14. 送測樣品送至核心設施後，多久可以拿到數據結果？
為能將樣品結果得到滿意的結果，本核心設施所有服務項目皆有一定流程及階段性，而送測樣品的完成速度，須視送測樣品量的多寡及樣品狀況而定。目前送測的樣品，從收件隔天開始計算3～7 個工作天內可將實驗數據給予送樣者。
 
15. 若登錄的樣品或送樣者資料有誤，可否代為更改？
個人資料與送樣資料，本核心設施人員皆無法進行更改，故建議重新填寫送樣資料。
 
16. 若實驗室主持人長期不在國內，申請單簽名處該怎麼辦？
請實驗室主持人以E-mail 方式(內容請載明指定之代理人姓名)寄予核心設施
(mass@ibms.sinica.edu.tw)，以玆證明代理人行使主持人權限。
 
17. 放射性、毒性樣品可否送件？
因考慮工作人員安全性問題，恕不受理放射性或具(生物)毒性樣品。
 
18. 撰寫論文時，由核心設施服務之數據應該如何敘述實驗方法？
使用者利用核心設施之服務進行論文發表時，建議之致謝撰寫方式請參考文件下載頁面之致謝說明及草稿。質譜分析撰寫可於文件下載頁面下載相關條件與方法之草稿。
1. Could I request the Core Facility to do pretreatment(Insolution/in geldigestion, etc.) of samples?
A: As the constraints on human resource, the core facility only provides the training and protocol downloaded for users. Beginners could see an online instructional video in training courses page, and the training courses will be held in March and September each year. The related information was
published in training courses page, please follow the instructions to registration.
 
2. What state should the samples be treated before submitting to the core facility?
A: Please cut the gel (be careful to avoid any contamination of keratin) and put in eppendorf, after that bring to Core Facility to do In gel digestion.
 
3. Should we need to prepare the reagents by ourselves when go to Core Facility to do experiments?
A: The Core Facility would supply all of the related reagents and consumables , please cherish resources and be careful to avoid contaminating the reagent.
 
4. Could the samples with silver stain be submitted to analyze?
A: The reagents of silver stain contain aldehyde that may affect the performance of mass spectrometry, so it is inappropriate to analyze by mass spectrometry. For the samples of low concentration, the Core Facility recommends to use the high sensitivity and fast SYPRO Ruby to stain. This stain method would not affect the performance of mass spectrometry.
 
5. How to estimate the amount of protein samples?
A: Please use an appropriate quantitative method to estimate protein amount, and inform us what quantitative method you used when submitting. The Core Facility could provide uBCA kit for estimating protein amount of insolution digestion samples. The samples of ingel digestion could run gel with a known amount of internal standard protein, such as BSA, to estimate the amount of protein after stain. Please note that the protein will have different degrees of loss after peptide extraction and desalt, so be careful to deal the subsequent procedure in order to get high recovery.
 
6. What are the notes of sample preparations for LCMS/MS?
A: Please refer the Notes in the submission page.
 
7. How to check the progress of sample analysis?
A: In principle, the Core Facility would follow the received order to analyze samples on machine. But the core facility has the right to adjust the order, according to the analysis capacity, sample character, and analysis time of individual sample. If there is any question for progress, please contact with Core Facility. (If the equipment was out of work or under maintenance, we would publish the information on the news page. The time for data return may need to extend.)
 
8. How to get the data?
A: After the analysis, the data will be sent to User and Principal Investigator by email. If the file is too large, the data will be uploaded to the FTP of your lab, and inform you by email or telephone to download data.
 
9. Could the leftover samples be taken back?
A: The leftover sample was 10%-50% of original amount. The Core Facility could keep the leftover samples for three months. If user would like to take back, please request within three months.
 
10. Would the Core Facility keep all of the data?
A: The original data would be kept in database of Core Facility for one year from the data being sent to the user. Because the storage capacity is limited, all of the data would be deleted once beyond the expiry date.
 
11. How to apply the service of relative quantitative analysis of protein expression?
A: Before to apply this service, users have to consult and discuss with Core Facility to make the reservation for the instrument and confirm the cost.
 
12. Could the users choose which instrument to analyze?
A: The Core Facility will choose the appropriate mass spectrometer to analyze for high quality results.
 
13. Who could I contact if have question for data?
A: If you have question for analysis result, please contact with Core Facility,we will explain the detail for you.
 
14. How long could I receive the data after the sample submission?
A: The time required for analysis of samples depends on the total amount of the submitted samples. In general, the users could be received the data within 5 workdays after submitting the samples.
 
15. If the information of sample or user is incorrect, could you change thatfor us?
A: The core facility staff can change neither user profile nor sample information, so we recommend refilling in the Service Request Form.
 
16. How to deal with the signature of the Service Request Form, if the principal investigator is longterm absence from Taiwan?
A: The principal investigator could send a message (please set out the name of assigned agent) to core facility by email(mass@ibms.sinica.edu.tw) to prove the competence of agent for principal investigator.
 
17. Could I submit a sample of radioactivity or toxicity?
A: In consideration of the safety of staff, the core facility doesn’t accept the radioactive or (cyto) toxic samples.
 
18. How do I describe the experimental methods for the data provided by core facility when writing papers?
A: When users utilized the data by core facility service to publish paper, about the written acknowledgement please refer to the acknowledgement statement and draft on Download page. And regarding the mass spectrometry analysis, the draft of related conditions and methods could be also downloaded on Download page.