Vo.5 No.1 - アジレント・テクノロジー株式会社
Contents
1. 200 2010 3 31 25 OFF Serum Plasma 1 2 3 4 5 678 9 kDa 200 116 97 66 1 55 a 2 37 3 31 4 a 5 22 6 14 7 8 6 9 1 HSA Hu 2 Hu 6 Hu 7 Hu 14 LC MARS2. Keynotes PURE system 69 Shimizu et al Nat Biotechnol 2001 1 RNA RNA DNA RNA 100 RNA 8 2lEe BB
3. 3 A B W O C RNA directed evolution RNA RNA
4. 2010 3 31 50 OFF FFPE 21 28 4 Zinc alpha 2 glycoprotein precursor Antigen K167 1 FFPE LC MS FFPE Protein Extraction Solutlion FFPE FFPE Protein Extraction Solution LC MS FFPE LC MS FFPE Protein Extraction Solution 3 FFPE 2
5. MS MS MS MS UIPIEF 1200 HPLC Chip MS Agilent 6200 Accurate Mass TOF MassHunter Workstation 1200 LC Agilent 6500 Accurate Mass 0 TOF MassHunter Spectrum Mill Agilent 6400 LC MS MassProfiler Professional HPLC Chip Phospho chip
6. IPG strip OFFGEL OFFGEL Fractionator Gel OFF 1 OFFGEL IPG strip 12 24well Frame Frame well IPG strip well IPG strip well 2 well Well
7. BSA 1 500 Pyrococcus furiosus Complex Proteomics Standard Pyrococcus furiosus ORF ftp ftD ncbi nih govV genbank genomes Bacteria Pyrococcus_furiosus Pyroc
8. Kita et al ChemBioChem 2008 Vol 9 3023 3028 Mx3005P PCR GFP RNA Kazuta et al Mol Cell STS 2008 1 UIDU IO 2
9. http Www symbio ist osaka u ac jD employmenUindex html 12 FAU gz P9 0 MAHS mRP HPLC LC MS HPLC AgilentZ0RBAX Agilent ZORBAX HPLC 2 1 Agilent ZORBAX
10. GFP 2 Mx3005P GFP 4 300 12 2 2 4 3 Tetracysteine promoter 1 ORF product Synthesis 37 C 3h GFP synthesis _ ES 2 GFP
11. MARS 14 3100 OFFGEL MassProfiler Professional 2 N Agilent 6400 LC MS MRM
12. 2 O1 Oo TY Agilent 6224 TOF MS CE TOF MS 2 MassHunter MFE Agilent G7100 Agilent G7100 Agilent 6224TOF CE TOFMS MassHunter MassHunter Professional 48 000 000 3 500 000 psorEoMlcs Agilent 1290 Infinity LC LC LC MS UHPLC Problem Solution Agilent 1290 Infinity LC N
13. A HPLC Agllent 1100 1200 HPLC HPLC LC Agilent HPLC A HPLC 5 2mm x 4 95mm 100uL HPLC 4 0A
14. 1 8um 800 HPLC 1 Agilent Z0RBAX HPLC A 1 ZORBAX HT RRHT Z0RBAX HT RRHT 2um 1 8um LC MS 2 1mm 4 6mm 2 600Bar 2 gt dummrr ro mm mhr SE Ct Lm 2 ZORBAX HT RRHT 1 8um RRHT 600Bar
15. GU5 saas 2S2Z0 Micro Total Analysis System uTAS Q A RNA RNA PURE system Keynotes 1 Keynotes 1
16. he Complex Proteomics Standard 7 Protein Extract Proteomics Standard 500ug Proteomics Grade Irypsin 100ug Proteomics Grade Trypsin Proteomics Grade Irypsin 100ug 400510 240310 5 sm Ng 20 2 Complex Proteomics Standard TPCK
17. Anti Albumin resin Hu 2 Anti lgG resin _ Anti Transferin resin Anti lgA resin Anti Haptoglobin resin a 1 Dsin resin Hu 7 Anti Fibrinogen resin J Anti a 2 macroglobulin resin Hu HSA Hu 2 Hu 6 Hu 7 Hu 14 1 2 6 7 14 Hu HSA 50 55 Hu 2 69 Hu 6 Hu 7 Hu 14 85 94 1 Mouse3 3 Albumin lgG Transferrin 98 99 20 200 uL Anti lgM resin Anti a 1 acid glycoprotein resin Anti Complement C3 resin Buffer A EN 0 DE i Anti Apolipoprotein A I resin Anti Apolipoprotein A II resin Anti Transthyretin resin Buffer B 8
18. 2 ZORBAX High Definition RRHD ZORBAX High Definition RRHD 1 8um Z0RBAX RRHD Agilent 1290 Infinity LC HPLC 1200 bar RHRHD 1 8um 50 100 150mm Solvent Saver HPLC Agilent ZORBAX Solvent Saver 3 0mm 3 0mm LC
19. Complex Proteomics Standard Proteomics Grade Trypsin Proteomics Grade Trypsin
20. Agilent 1290 Infinity 1 7um 150mm 100mm 2 BSA 567 5 ml min ET ET LL I 1 PF 2 Agilent 1290 1 7um 100mm 150mm 567 Agilent Q TOF MassProfiler Professional Agilent 1290 Infinity LC Ultra High Definition TOF Agilent G6540 Q TOF 2 Q TOF 3 MM MFIEI LM ll Ma 3 Agilent 1290 6540 0 TOF
21. SCX 2 3 HPLC OFFGEL Fractionator 4 LC MS SCX OFFGEL Gea hy Agilent 3100 OFFGEL G3100AA 2 348 000 Fractionator pH pl electrode 2 2 3 Protein A Protein B TPG gel pH gradient 2 OFFGEL Fractionator b Starter Kit Low Resolution Kit pH3 10 High Resolution Kit pH3 10 Low Resolution Kit pH4 High Resolution Kit pH4 pH 3 10 12well 12 188 6425 188 642Z4 188 642 188 6426 188 6444 149 000 149 000 149 000 149
22. 3 Co 4 6mm 1 2 1mL min 1 2 0 43mL min 1 2 3 4 3L ZORBAX Eclipse Plus C18 Solyent Saver 3 0 x 150mm 3 5 m 40C 0 19 41 9 UV 269nm 3 Agilent HPLC 5069 3635 DEAE S03 Protein A 5069 3636 5069 363 5069 3639 mm A QA Agilent HPLC A Agllent HPLC lgG lgM DNA
23. HPLC Agilent HPLC LC MS 1 1 http WwWw chem agilent com 1 HPLC LC MS AA AM NM 0 HPLC A HPLC 0DS HPLC LCMS HPLC Agilent Z0RBAX HPLC A Agilent ZORBAX HPLC US NEWPOHT Agilent HPLC
24. Q A vitro Q PR 21 10 ERATO
25. Agilent HPLC Chip 6500 Accurate Mass 0 TOF Ka MS MS SALSA Spectrum Mill MassHunter BioConfirmation HPLC Chip HPLC Chip 6500 Accurate Mass O TOF
26. Agilent 1200 HPLC Chip Agilent 6000 LC MS in solution digestion in gel digestion 2010 3 31 50 OFF Counts X 105 Acquisition Time min 1 SN GEFENARDEE TANGERISEY PEPPER PALKPLALVY NGHSENVEYD DSODKAVLKD GPLTGT 1 1 1 1 fi 120 JNES TAAOOPDGLA YWGVRMSD RBACIKYEP TDS STDEFPNEDPG 0 SLLPNVLDYW TYPGSLTTPP LLESVTWTVL AEGEPELLML YRLV QFHFHWGSSD DQGSEHTVDR KKYAAELHLV 240 RE WRNRGERR carbonic anhydrase I Proteomics Grade Trypsin Agilent 6520 Accurate Mass Q TOF HPLC Chip 89 9S999 49 000 4 900
27. QuikChange Lightning Multi Site Directed Mutagenesis Kit p7 1 DNA 5 kb QuikChange Lightning Multi QuikChange Multi kit Cn Muti 2 DNA 1 1
28. 50 mRP C18 _ mRP C18 LC MS MS 60 0 mRP C18 LC LC 4 mRP C18 _ MS 10 mRP 400
29. www agilent com chem jp 0120 477 111 PpROTEOMICS mm Problem Solution Mulitiple AU Removal System MARS 8 MARS MARS LC LC HPLC
30. Agilent G6224AA TOF CE MS CE CE MS CE MS Agilent CE MS Single Quad TOF Q TOF LC MS CE CE MS 1
31. pH 2 13 IqM DEAE pH 3 9 DNA S03 pH 2 13 A IgG 3 IgG A lgG 141 000 41 000 41 000 160 000 5 2 x 4 95 5 2 x 4 95 5 2 x 4 95 5 2 x 4 95 14
32. FF fi THE r 1 hl 3 GFP Q Mx3005P PCR 4 3 Mx3005P Mx3005P Mx3005P 1 180 1 4 Mx3005P PCR
33. Fusion Pf DNA DNA Dpn 3 SR DNA Dpn DNA Kit XL10 Gold Ultracompetent Cells 3 3 50 1 95 DNA QuikChange Lightning Multi Enzyme Ble
34. N N 250 bar 4000 ps pH lt 5 80C pH 5 8 40C mRP C18 4 6 x 50mm 5188 5231 99 999 75 000 mRP C18 0 5 x 100 mm 5188 6510 99 999 5 000 2 1 x 75 mm 188 651 1 99 999 75 000 10 NNE EE NN EE eg Vocat Lead Vocall Vocalist Vocalist Vocalist
35. Protein Sample Prep Kits FFPE Protein Extraction Solution 25 1 25 ml FFPE Protein Extraction Solution 100 4x1 25 ml PPS Silent Surfactant 5 x mg PPS Silent Surfactant 10 mg 52 0099 26 000 93 000 18 000 27 500 400925 400926 400500 400501 36999 SSr999 PROTEOMICS HH HPICO 5 48 OFFGEL Fractionato Problem HPLC Solution OFFGEL Fractionator IPG strip IPG strip
36. 1 ug 96 well ELISA Proteins ldentified Mammalian Proteins 31 34 8 Cancer Related Protein Agilent FFPE Protein Extraction Solution 89 Vertebrate Proteins 33 37 26 35 1 Periostin precursor 90 K167 Macrophage migration inhihitory factor PPS Silent Surfactant PPS Silent Surfactant pH
37. Problem 2 8W NS LC MS FFPE LC MS FFPE FFPE FFPE Protein Extraction Solution 1 1 Total Yield pg 1 4
38. QuikChange Lightning Multi Site Directed Mutagenesis Kit 210515 210513 210515 210513 210516 210514 10 30 10 30 QuikChange Lightning Multi Site Directed Mutagenesis Kit Academic 216 000 172 000 p QuikChange Lightning Multi Site Directed Mutagenesis Kit Commercial 420 000 Problem Complex Proteomics Standard Pyrococcus furiosus
39. 449 999 330 000 783 889 527 250 3 439 999 2 354 250 388 999 291 000 65 999 488 250 2926 999 2 194 500 295 999 221 250 496 899 372 000 18 099 135 750 68 008 126 000 369 999 276 750 620 999 465 000 2 86 999 2 089 500 295 99 496 G98 25 999 188 250 194 999 145 500 48 699 111 000 76999 57 000 68 999 138 750 PpROTEOMICS mRP C18 Problem Solution HPLC mRP Macroporous Reverse Phase Column C18 95 pH pH
40. micro wells filled with buffer pH p 1 O electrode f protein 5 pH gradient 1 OFFGEL Fractionator a hh Starts with micro wells filled with sample diluted in buffer electrode 1 pH gradient hy pH pl pH pl Protein B m electrode Protein A 1 Well Focusing 3 NUT ll ll Ml inl bbl 3 OFFGEL Fraction 3 SDS PAGE OFFGEL Fractionator 4
41. 401449 50 0FF 2010 3 31 Mx3005P Heal lime PCR System 400510 Complex Proteomlcs Standard 204310 Proteomlcs Grade Trypsin 400500 PPS Silent Surfactant 400501 PPS Silent Surfactant 400925 FFPE Protein Extraction Solution 400926 FFPE Protein Extraction Solution 25 0FF 2010 3 31 H091030 2 LC MARS Newl High Capacity MARS YI iM High Capacity mRP C18 mRP C18 4 6 X 50 mm Hu 14 4 6 X 100 mm Hu 14 10 x 100 mm Hu 14 4 6 X 50 mm Hu 4 6 Xx 100 mm Hu 10 x 100 mm Hu 4 6 Xx 50 mm Hu 6 4 6 x 100 mm Hu 6 4 6 X 50 mm Hu 2 4 6 X 50 mm H
42. Proprietary IgG Agilent 1290 Infinity LC Agilent 1290 Infinity LC Agilent 6500 Q TOF Agilent MassHunter MassProfiler Professional 4 mr OF i mm FT FN mT I EE PN RE FH FE PT PN FE FH FE I mS LE 4 0 TOF Agilent 1290 Infinity LC 11 000 000 47 500 000 O IGP lt EAA 3 3100 OFFGEL Fractionator MARS mRP NN
43. mRP HPLC A OA OE UO Et C18 NM 1 HIN A _ iN hi i HH LC Yi mRP C18 1 MARS 50 95 99 2010 3 31 EE NUDE 25 OFF Proteins Identifed Human Serum 40 Immunodepleted 181 a Human Se
44. High Capacity MARS MARS High Capacity 4 6 x 50 mm Hu 14 4 6 x 100 mm Hu 14 10 x 100 mm Hu 14 4 6 x 50 mm Hu 4 6 x 100 mm Hu 10 x 100 mm Hu 4 6 x 50 mm Hu 6 4 6 x 100 mm Hu 6 4 6 x 50 mm Hu 2 4 6 x 50 mm Hu HSA 4 6 x 50 mm Hu 6HC 4 6 x 100 mm Hu 6HC 10 x 100 mm Hu 6HC 4 6 x 50 mm Ms 3 4 6 x 100 mm Ms 3 0 45 mL Hu 14 0 45 mL Hu 0 45 mL Hu 6 0 45 mL Hu 2 0 45 mL Hu HSA 0 45 mL Hu 6HC 0 45 mL Ms 3 2 SDS PAGE 188 655 188 6558 188 6559 188 6409 188 6410 5188 641 1 185 5984 185 5985 5188 8826 5188 6562 5188 5332 5188 5333 DIGGHO936 5188 521 188 5218 188 6560 188 6408 188 5230 5188 8825 5188 5334 5188 5341 188 5269
45. CE MS 1 144444MIII4 U4MH11 I Buffer 1M formic acid Sample CysCys 62 5mmol 250 mmol Caplllary fused silica L 100 cm id 50 mm Injection 150 mar 1 Agilent CE MS MS CE TOFMS MassHunter Agilent G6224 TOF MS NN 1 WE RR EF SN Scan 40Hz 6 MassHunter Molecular Feature Extraction MFE
46. 000 149 000 pH 3 10 24well 12 pH 4 7 12well 12 pH 4 7 24well 12 4 3 12 METABOLOMICS ew Agilent CE TOF MS Problem LC MS 1 UV Vis Solution Agilent G7100 Agilent G6224AA TOF MS CE MS
47. 229 IN 1 PURE system 69 RNA RNA QB IN MG 9 RNA RNA RNA MC OR TE 1 RNA RNA RNA RNA HNA PURE system RINA HNA 1 Q promoter T5 tag Mx3005P PCR PCR
48. 0 330 000 527 250 2 354 250 291 000 488 250 2 194 500 221 250 372 000 135 750 126 000 276 750 465 000 2 089 500 221 250 372 000 188 250 145 500 111 000 57 000 51 000 138 750 111 000 75 000 75 000 75 000 8 F
49. 1 cm2 10 um PROTEOMICS Solution FFPE Protein Extraction Solution FFPE LC MS PPS Silent Surfactant LC MS FFPE Protein Extraction Solution 2 Epitope Preservation liver 0D 450 nm Agilent R actin GAPDH Supplier 1 Antigens 2 B actin GAPDH 1 cm2 10 um FFPE FFPE Protein Extraction Solution
50. StrategieS VOLUME 5 Index LC MS 1 2 LC MS 2 HPLC 0FFGEL Fractionaor Agilent CE TOF MS Agilent 1290 Infinity LC 6 4 m4 UHPLE 7 8 new 5 new ew new gt CO No On new mRP C18 new 2009 2010 13 14 15 Agi
51. e al UIMIO CS MO 2 6 UHPLC UHPLC HPLC UV 0 Uee 2K UN ES Agilent 1290 Infinity LC Agilent 1290 Infinity LC 120MPa UHPLC 2um 100mm 1 Agilent 1290 Infinity bar 1200 1290 Infinity 1000 A 800 D CT 600 400 200 0 1 2 3 4 1 Agilent 1290 Infinity LC
52. lent Technologies MUTAGENESIS Problem 5 QuikChange Multi Site Directed Mutagenesis Kit 3 QuikChange Lightning Multi Site Directed Mutagenesis Kit DNA 5 Solution QuikChange Lightning Multi Site Directed Mutagenesis Kit 50 p 1 SEED CE ESE TN TE 3
53. nd OK LU u LT lt Dpn Dpn I XL10 Gold Ultracompetent Cells 1 QuikChange Lightning Multi Site Directed Mutagenesis Kit QuikChange Lightning Multi Site Directed Mutagenesis Kit 4 Reaction Time Comparison for Incorporation of 4 Point Mutations Using OuikChange Lightning Multi vs Original 0uikChange Multi 0 60 120 180 240 300 360 420 480 540 600 660 4 kb 5 kb 6 kb 6 kb Minutes 0uikChange Lightning 0uikChange Multi 2 QuikChange Multi Lightning QuikChange Multi QuikChange Lightning Multi Site Directed Mutagenesis Kit 210516 210514
54. occus furiosus LC MS 6JIR62I Solution Complex Proteomics Standard Pyrococcus furiosus 1 500 Complex Proteomics Standard
55. rum E 380 ug Fraction 16 71 oo Fraction 22 97 Fraction 28 99 Sm 8 99 Combined Fractions 394 16 22 28 32 HPLC Human Serum data collected off line by 2D LC MS MS Ua with the Agilent Multiple Affinity 2 pH 0 9 8 0 6 0 pH PEEK HPLC 1 16 10 32 PEEK 2 um C18 5 um
56. u HSA 4 6 Xx 50 mm Hu 6HC 4 6 x 100 mm Hu 6HC 10 x 100 mm Hu 6HC 4 6 X 50 mm Ms 3 4 6 x 100 mm Ms 3 0 45 mL Hu 14 0 45 mL Hu 0 45 mL Hu 6 0 45 mL Hu 2 0 45 mL Hu HSA 0 45 mL Hu 6HC 0 45 mL Ms 3 46 Xx 50 mm 0 5 x 100 mm 2 1 X 75 mm 500 hd 100 pg 5 X 1 md 10 mg 25 100 0188 6557 0188 6558 0100 6559 0188 6409 5166 6410 5166 6411 5105 5904 0105 5965 5100 0026 5160 6562 9188 5332 0188 5333 0188 5336 0188 5217 0188 5218 0188 6560 5160 6406 5160 9230 0100 0025 9188 5334 5160 5J4 1 0100 9269 0188 5231 5166 6510 5160 6511 T 192 8510 9 1 eg 0120 477 111 emall japanQagilent com WWWw agllent com chem jp Mx3005P Real Time QPCR System 500 066 446 600 793 099 19 900 S96 990 65 990 926 000 295 990 496 990 6 890 66 990 369 860 626 990 796 990 295 660 496 990 ZS5F090 19 6600 46 099 76 660 69 660 65 9090 48 990 3 400 000 49 000 4 900 18 000 27 500 26 000 93 00
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