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Peptide Mass Fingerprinting 分析手冊 ( 使用前請洽公用儀器室 )

Peptide Mass Fingerprinting 分析手冊 ( 使用前請洽公用儀器室 ). 3. 選取欲分析的圖 檔. 2. 開啟檔案. 1. 由此進入 Data Explorer 分析 ( 本範例以 BSA 為例 ). 4. 欲分析的 Mass 圖譜. 9. 選取適當分子量範圍去除雜訊 , 選 Use Advance Settings. 5. 基線校正. 7. 背景扣除. 6. 選 OK ( 參數設定不動 ). 8. 選 OK ( 參數設定不動 ). 10. 增加質量範圍段落數目. 11. 刪除質量範圍段落數目.

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Peptide Mass Fingerprinting 分析手冊 ( 使用前請洽公用儀器室 )

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  1. Peptide Mass Fingerprinting分析手冊 (使用前請洽公用儀器室)

  2. 3.選取欲分析的圖 檔 2.開啟檔案 1.由此進入Data Explorer分析(本範例以BSA為例)

  3. 4.欲分析的Mass圖譜

  4. 9.選取適當分子量範圍去除雜訊,選Use Advance Settings 5.基線校正 7.背景扣除 6.選OK (參數設定不動) 8.選OK (參數設定不動)

  5. 10.增加質量範圍段落數目 11.刪除質量範圍段落數目

  6. 12.選Range1,質量範圍799-900,選OK 13.把小分子量去除,在%BP Intensity輸入一個適當的值(圖中的虛線),濾除雜訊 左鍵拉開質量799-900,右鍵可選Single Unzoom或Full Unzoom

  7. 14.選Range2,質量範圍900-1200選OK,接著處理其他質量範圍至400114.選Range2,質量範圍900-1200選OK,接著處理其他質量範圍至4001

  8. 15.Range3,質量1200-1600 16.Range4,質量1600-2000

  9. 17.Range5,質量2000-2400 18.Range6,質量2400-4001 19.選確定

  10. 去除雜訊後的Mass圖譜(雜訊不被標示)

  11. 20.Peak Deisotoping,選OK(參數設定不動) 做Peak Deisotoping後的分子量Mass圖譜

  12. 21.Copy Mass List準備記錄與搜尋 22.開啟Protein Prospector網站(http://prospector.ucsf.edu/) 23.由此進入

  13. 24.選擇Database(常用的有 Swiss Port,NCBI nr,Genpept) Cysteine modification 樣品名稱 參數設法以Gel上決定的分子量+10K為準,可自行調整

  14. 25.將剛才在Data Explorer中複製的Mass List貼入後(滑鼠右鍵,全選,貼上)按Start Search 要記得哦!

  15. 26.Database search 常用的有 Swiss Prot, NCBI nr, Genpept,search完印出後,可按Back(上一頁)回去更改Database再serch一遍

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