2次元電気泳動のコツ
Contents
1. WS X pH Y2. A F Gc FA EE lt _ DTA free 2CHAPS 5 ITLEOT A 100 3 pH8 8 9 0
3. CSH AZc ae Michae M S mm ms H 1 Protein SH or Proteih S G G G H Hz H 4 Protein SH FroteIh H NHs AITO 2 D A IEF B
4. EzStain Silver EzStain Silver 16 000 2D 2 CBB 10 20 20 40 rR Dar SA 30 60 27 1 mm 28 1 CBB EzStain AQua EzStain AQua 9 800 ATTO
5. 2 2 1 2 SDS PAGE En a F i Mi a 2D 2D 2 lt S N 0 1 2
6. 6 20 SR 26 25mM 192mM 0 1 SDS 1410 Run PAGE 35min 3 20mA 1 40mA 2 PAGE 76 000 78 000 CBB C B B
7. 24 2 2 y c 6 he 15 000 17 000 e 13 800 2D 2 1 2 3mp9
8. 7 8 2 D 0uant Kit 9 DTT C SH CS S Michae Michael 2
9. 1 UI 1 50 nuL 1 5 10 6
10. 2 SDS 1 1 2 1 1 2 2 100 uL 2 2 2031 2 U 1 2 J 25 1 SDS PAGE Im
11. 8 80 2 oo 4 2 2D 2 es 1 pH pH 3 10 pH 3 8 2 D 1 pH 2 12 5 60X60X0 75mm CBB 2 NT rT H 8 at Ey rr ii er ER IT 5 a k 1 N 8 5 id 0 1 2 he CR Wa le ED i TH Ny at Wh nt Ed NT IPG 2
12. 80 Tk 2 TEE 5 EzApply2D Kit 0 000 1 2 EDTA free EDTA 3 CHAPS TritonX 100 4 pH8 8 9 0 9 5 DTT CS S DTT 20 1
13. 90 000 2 1 native lt 2 D 2 75mm 15 50 uL a oH gt NN pH
14. 5 50mm 5mm ____ 2 00 _ 10mm 1 13 2D 2 1
15. lt 2 CBB 50mm 100 ug 75mm 200 ug 2 50mm 1 lt 2 ug 75mm 2 4ug 10 unL 3 0 0V 50m 150 75mm 210 1 mA
16. pH 8 8 lt 90 80 i 2 50mm 100 u g 75mm 200 u gs 1 70 300V 50mm
17. XX Y2 5 0 0 MS MS gt a co 1 nn 1 400 ldentification result Protein list Protein detail Protein sequence Selected protein Rank 1 Expectation 0 002 Covera ge 20 9p S50 Mass 32 26 kDa Matched peptides 5 Measured peptides Protein information 13435578lgbIAaH04705 1AaH04705 BCOO47O06 heterogeneous nuclear ribonucleoprotein C Mus musculus Matched peptides Meas mass Ave pic Comp mass 886 365 Mono 885 400 IMYSYPaR 942 519 Mono 942 566 OvPPPPPIAR 1328 652 Mano 1328 648 1 IiFAFVHYVNER 1681 83
18. p8 25 50m_ A lt A 100C 20 A B 2 50mm 75mm
19. DTT 5 2 4 10C 1 10 50mM 50 000rpom 4 20 6 1 10 1M 1M CS 6 15 000rpm 4C 30 60
20. 5 10 3 lt 4 9 000 10 9 500 10 2D 2 9 p 10 11 NN OK 12
21. EDTA free CHAPS Tritan 100 p 20 C DTT 1 3 CSH SS it HOOC C C SH aa 1 NHH S S 2
22. 1 2 lt DTT DTT 20 1 pH 88 90
23. 2 D 70 ug 1 2 5x75mm 3 10 2 12 5 0X80X 1mm C BB Fah BEhLEm Fe Eh 0 es a F Ps 3 kEm gt ATTO0 2D 1 A I PG 1 IPG 2 10 TEL 03 3814 4861 Fax 03 3814 4868 E mail info atto co jp 113 0034 1 5 32 2D ee 4m 2 2009 4 1 ES FV fTITTD A 113 0O34 1 5 32 3 3814 4961
24. 1 1 pH S S etc 1 2 1 2 1 1 6 lt 1
25. 1 2 1 1 2 eg _ a i BPB 3 5m PAGE 3 5 8 0 9 0 y ZZj ge 6 l
26. 150 5mm 210 1 2 1 2 2 1 J C B B CB B SDS PAGE C 1
27. 2 1 1 2 2D 20 100 14 300 i e 1 pH5 8 2 12 5 2 1OI 9 39NIOSOI OLIV AOO1ONHO LOI Y 3 9N IOSOI OLIV OO0 1ONHO LOI 9 9N IOSOI OLLV AO01ONHO LOId ON IOSOI OLIV AOO1ONHoO LOI 9 3 ON IOSOI OLLV AOO01ONHO LOI 9N IOSOI OLLV 5O 1ONHO LOI 9 39N IOSOI OLLV 1 2 2 0 2
28. 03 3814 48BH 3 3814794 03 31 4858 110 0016 5 91 6 D3 5818 75B0 03 5818 7553 53U UU54 1 U6 63B 7 1g 0B 63B5 71 URL http www atto co jp e mail infoQatto co jp e mail osaka atto co jp
29. 10mM 40mM DL 14 6H gt 200mMN a OH CH2 10M 40mMDL L pH 16 lt 0 01 BPB BB 17 0 Farrell pH overnigshD Vr 300Vx210min 70Vx900min 15hr
30. 5K 3 1 3 2 ro 19 2 2 SDS 2 100mL 10 18 TCA
31. 19 9 20 1 2 gt 3 21 50mM Tris Cl pH6 6 2 6SDS 0 0 1 BPB 2D 2 3 1 X3 120 SDS 10 ATTO BIOSCIENCE amp BIOTECHNOLOGY ATTO BIOSCIENCE amp BIO
32. 2 2 agagrG7 1D E sc qdyva sas az A rg J a eT PAGE ATTO Corporation ATTO Corporation Osaka 1 5 32 Yushima Bunkyo ku Tokyo 113 0034 2 1 7 Minami Morimachi Kita ku Osaka City TEL 03 3814 4861 FAX 03 3814 4868 T 530 0054 URL http www atto co jp TEL 06 6365 7121 FAX 06 6365 7125 2D 2 EAC Ss 2 2 2
33. BIOSCIENCE amp BIOTECHNOLOGY ATTO BIOSCIENCE amp BIOTECHNOLOGY ATTO BIOSCIENCE amp BIOTECHNOLOGY ATTO BIOSCIENCE amp BIOTECHNOLOGY ATTO BIOSC 2
34. Mono iMIAGHVLDINLAAEPK 2100 728 Man 0 2D 1D pH5 8 2D 12 5 293 40ug heterogeneousnuclearribonucleoproteinC 1681 305 2100 870 SAAEMYGSSFDLDYDFOR 2 1 600 000 2D 2 1 2 lt DTT DTT 20 1
35. TECHNOLOGY ATTO BIOSCIENCE amp BIOTECHNOLOGY ATTO BIOSCIENCE amp BIOTECHNOLOGY ATTO BIOSC SDS PAGE Laemmli 309 6 CH8 8 pH6 8 1096 APS IEMED APS TEMED 4 0 1 2 0 22 5 20 10000400000Da 23
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