“WAKO BIO WINDOW” No.69(2005.08)
Contents
1. 10000 0000 20000000 00000 00000 0 OUOU 2m 0000000 Im 0000 0028309005 3 5 5 0 000 0 0000000 000000000000 2 0000 00006 0000 2 00000 00042921 0 000 00000 28 0 2 3 00000 S6100 00000 00 94737 0 00000 00000000 00090 _____ 500 __ 9610 0 00000 500 0 200 0 00 0010 5001 0 0 50407 25w 125m 5 25471 129 96500 00000 96100 0000 24 0000000 0 0 0 002839 18 247 000 0000000000 0 000000 0 0 0 00 249 00 00 0000 000 00000 00037 21 0 0 00000 0 510p 4 00000 00028 3092 3 0000 00 000000 0000000 Sc Transformation Reagent 201 O0 OOOO DNA Hg 21 0000A L0 L 0000 20 0 BRUNE 6100 0000000000000 02. HHAH 0 esterase F 2 190 22 o 04 11 Maeda H Fukuyasu Y Yoshida S Fukuda M Saeki K Matsuno H Yamauchi Y Yoshida K Hirata and Miyamoto K Angew Chem Int Ed 43 2389 2004 2 Maeda H Matsuura S Nishida M Senba T Yamauchi Y and H Chem Pharm Bull 49 294 2001 ooono o n 00000000 029 15381 BES H202 25 000 K W Wako Bio Window AUG 2005 No 69 DNAD 100000 5 0 0 0 000000000000 0 Pore 1 00000 00 0000 0000000000000 5 00 controlled Glass 0000 0 00000 0 0 0 0 000 0 00000000 First Nucleotide dN Genogiass T 0 0 0 0 000000000 y Amino 0 000000000 10000000
3. CeBND 0000000600 5 0 0 0 0 0 0 0000000 000 0 0 00000 0 0 0 0000 0 0 0 0 00 J U UUUUUUUUUUUUUUUUUUUUL 7 540 8605 0 00 00000 0 0 00 1020 2 0606203 17881 0000 103 0023 000 00000 00 0 00050 130 2 0303270 82431 0000 00000 3 0921622 1005 00 e0000 0 082 285 6380 00 3 052072 078800 04576 20610 0 O 3 029858 227800 e0000 0 280221222 3070 00 000000 01171 0289 0 O 0000010 0 O 0120 052 099 1000000 0O O 0120 052 806 DM 000 0 0 0 0 0 00 0 biowin URL http www wako chem co jp 05 219 850 1 DN
4. 000 582 84111 71009 3 588 84113 71009 4 YPS 0000001 35800 0000 0000000000 1 293 0 0 000 0 0 60 00 00000 2 5 000 0000000 ________ _______ 554 73421 71183 3 0 000 0 0 000 33 800 U N 0 00000000 0 Biolegend 0 0 0 2000 123 00000 0000 0 00008 Wako Bio Window 00000 0 78 11527 Ba m j e __ Wako Bio E mail biowin weko F A X 06 6201 5964 Wako Bio Window AUG 2005 No 69 210 000000 0000 000 0 0 0 DL O0 0 0
5. 800 14 99 00000000 0000 123456789 10111213 14 1516 M ODE 0 0 QuickGene 800 2 30 260 280 1 000M00000 0g 000 00010 80 QuickGeneg 0000000 S QuickGene 800 193 00090160 A0000 191 10000 00000 000 00 ication Guide No 1 No 10 00 00 00 0 00000 00000 No 11 000 60 No 20 000000000 0 0 0 O 0000 DNA 81 81111 No 31 0 0 0 0 0 293 48 0 0 0000000 O 130 8100 00000000 DNAD 00 00000 0 0 0 0000 DNAD 131 81000 000000000 0 00000 130 8100 0 0 0000000 9 0000000000 00 000000000 Svp 0000000000 DNAD 0 0 0 0 000 0 61 8110 a 00000000 00 80008 0000000000 000 0000000 _________ o 636 05601 __ QuickGene 300 H 10000 633 05611 __ 5 0005 __________ 9010 3w 630 05621 51 400
6. gt 0000 0 0 0 0 0 0 0 0 0 0 0 0 0 000 000 0 000 D0 00000000 29464001 DsDD cDNA Subtraction Kit Wako 1 WOD 98000 0000D L 0000010 574 98041 Polymeras amp Fidelity Systems 0 07 016 25 000 290 61501 14 000 580 69991 77 400 294 61401 15 000 043 29291 dNTP Mixture __ _ ___________ _ 900 311 90151 Phenol Chlorofomy Isoamyi Alcohol 2220 2501 15000 052 07221 Ethan 95 _____ __ y y y O ___ _____ 80 546 01651 100bpDNA Step Ladd 100 150 ____ 312 01193 AgaroseS ___ ____ _________ 20 _ 1200 313 90085 50x ___ _______________ 900 314 90261 2 500 315 90061 EthidumBromideSolution __ 6 __ __ ___ 900 316 90101 Distilled Water Deionized Sterile ___ _ 400 000000 Zhulidov A Bogdanova A S etal D A Nucleic Acids Res 18 32 3 37 2004 Wako Bio Window AUG 2005 No 69 5 cerevisiad 00000 0600000 000 000 200 Saccharomyces cerevisiae 1
7. 55 000000000 000000 amg 0 600 0 gmon D00No _______ 00000000 Se res P ooy 0 999 Amino 75 29 00 0 o aA 1 0 Sodas 0 Spes Sas Sro 0 0 809729 00A Se mees 0A 0 2 000 0000000000000000 000000000 0000000000000 O 00000000 200500000000000 000000000000000 0 0000000000000 0 0 0 00 0 0 0 0 0 0 00 00 119 2710001311 19 16 45 0017191 29100011 51 16 45 FAXO 0 00 0 000000 0 000 0 600 00 0 1000 0 0 0000 0000000000 http www wako chem co jp siyaku info life article amaxa _sem2005 hm p00 Wako Bio Window AUG 2005 No 69 6 7 High Throughput Western Bio 32 0 0 0 High Throughput Weste HTW Bio 0
8. 4 0 0 0 0 000 00000 00 00000 000 OO 0000000 00000 00 90 OOU 013 20251 Apolipoprotein E2 Human recombinant 000000 010 20261 Apolipoprotein Human recombinant 000000 017 20271 Apolipoprotein E4 Human recombinant 000000 0000 00 000000000000000000 0 000000000000 00000 000 000000 00000000 5 16X 000 35901 890 16000 000 RTICES 96X 1200301 96700000 000 0 0 0 0 0000 0 000 GPCR 0000 0 000 00 000 ___ ___ 0000 ____ oromi 581 84821 5 160 0 0 0 1600 6 _ 8900000 588 84831 5 960 000 9610 0 H 9300000 583 84881 RT CES 1690 0 00 16100 0000 55 000 580 84891 RT CES 96X 9610 0 0000 120 000 Wako Bio Window AUG 2005 No 69 gt SCRC
9. OOOO gt Antibody Capture Affinity Ligand gt Wash Buffer 10X gt Non denaturing Elution 4 gt Denaturing Elution 1 gt Spin Columns gt Capture Tubes 000000 000000000000 2000 8 15 30 4 00200 0000000 30100 0 0 0 0 0000 0 0 000000 0 000 00000 0000 0 Protein 28 Host zs AFi 45 1 TEE za 2 41 Hi 22 ES ni 42 04 700 za 1 45 anj 220 05 716 1 05 M00000 Reg 00 0 O0 00000000000 00000000000 Misie Ta Myd 00000000 0 000000000000 OOUDOUN 000000000 Catch amp Release vs Helaj 0 0 O edk21 906 505 or 0 M0 L0 0 M0 L 00000000 525 0 000 00000 0 0 0 0 0000 1 Flowthrough 0 0 2 wesh 1 0 0 3 wash 2 1 0 0 4 wesh 3 0 0 5 1 6 edution2 0 0 0 7 edution3 0
10. 16 6 O 0000000000 0 p 13 Transforming Growth 1 Human Insect Cells recombinant p 15 Connective Tissue Growth Factor Human 15 15 16 Miser Antibody Extender Solution p 17 Catch and Release v2 0 Reversible Immunoprecipitation System p 18 R amp D Human VEGF C Quantikine ELISA 21 0000 _ 12 Calbiochem 22 p 22 0000 IFILM QuickGenel 800 p 10 80 p 11 DsDD cDNA Subtraction Kit Wako 2 DsDD cDNA Subtraction Wako p 3 S cerevisiad 10 00000 00 00 00000 00000 0 v 4 Step Ladder EE E E 6 Step Ladder E EE E E
11. Tester 00000 00 Driver 0 0000000000000 0000000000000000000 0 0 0 0 0 0 000 0 0 0 0500 cDNA Subtraction Library 00000 000 000 00000 00002 0 01 cDNA Library Subtracted 000 Tester 0000 00 Duplex specific nuclease 0000 0000 5 000 0 00 0 00 Duplex specific nuclease 0 0 0 0000 0000 2 7 sem EE yras ira Tester cDNA Library 1 Driver cDNA Library O 00 00 Testei ean 00000000 cDNA Library 0 00000 1 Tester cDNAQ a 4 000 50000 000 0 00 000 00 T T E Air 2 Lambda 210 5 00000 00 5 9300000 00000 000 000 0 00 H 0000 68 161 20 Tester cDNA Driver 0 2001 0 0 0 Driver cDNA 000000000 Tester 1000 00000 DSN D 21 0 0 0 00000 0
12. 00000 55176231 1700 __ ______________ 5330 D00000 000 0000000 ________ 0000 BVD 10 0 00000 000 00 0 000 0000 00 00005 2177 1486 GR 23118 NPY Y4 0 000000000 0 NPY Y4 0 0000 00 00000 0000 00 500p g i 00000000000000 0 00 0000 0000 50 mg 155 300 _ Neuropeptide human rat 200p g 29 300 000 000000000000 _ Neuropeptide porcine 2004 g 33 800 000 0 000000000000 _ 1177 Neuropeptide 13 36 porcine 500p g 40 100 NPY Y2 0 0000000 1176 Leu Pro Neuropeptide human 20049 27 100 NPY Y0 O 0000000 E Pancreatic Polypeptide human NPY gt Ala Aib Polypeptide 0000 NPY Y4 0 0000000 Wako Bio Window AUG 2005 No 69 000000 000000000 Human VEGF C ELISA Vascular Endothelial Growth Factori 00000000 0 00 000 00 00 0 0 00000 0 00 00 00 0 0 0 0 000 1 1 1 1 1 Chilov
13. 0 00 0 0 0 0 00 0 2 gt Dye Solution Lane 1 Step Ladder 500 10 0006 1500 1 000 2 000 4 000 10 000bp Lane 2 Step Ladder 100 5 0000 1100 400 850 2 000 5 0000 Lane 3 Step Ladder 50 1 5006 50 200 400 850 1 5000 Step 100 1 0006 00000 gt Step 100 1 0006 PY 888888 8 58 888 Lane 1 RNA Step 1 100 1 0006 100 200 400 600 800 1 000b Lane 2 in vitro D 0 220b 2 Formaldehyde Agarose Native Agarose 000 No 81 71 292 61201 DNA Step Ladder 500 10 0006 15 000 296 61101 DNA Step Ladder 100 5 000bp 15 000 290 61001 DNA Step Ladder 50 1 500bp 15 000 298 61301 RNA Step Ladder 100 1 0000 0 04m x 5 20 000 O Wako B S H207 3 O Acethyl 6 O pentafluorobenzenesulfonyl 2 7 1
14. 0000 000 0000 00000440 0000 0 0 0000 0000 00000 0000 Attoglow Westem Blot Analysis Ki 0000000 K3171250 1 0 0 O 0 0 0O 00 0X0 000000 000 0000000 ________ 0 High Throughput Total Protein Westem Blot Human Normal 320000 160000 I T Wako Bio Window AUG 2005 No 69 0 000000000000000000000000000000000 Frozen Tissue Array BioChanj 1 00 20 e RIU 0 00000000 0 0 2mm e000000000000000000 Pancreas Thyroid e E Cerebrum Cerebellum Lung F 7 Breast 4 8
15. 2 E EERTE EE SLET Skin Skeletal Muscle Kidney Heart e Stomach Small Intestine Liver Salivary Gland 40 01000 Frozen Tissue 20 0 0000 4 0 00000000 0 00 00000 0000 000 0000000 T6234700 2 Tissue Array H Aduit Normal Ti 40 specimens 162347005 rozen i Issue uman Issug speci 3 76235700 2 Frozen Tissue Array Human Tumor amp Normal 40 specimens 10 I T GUC 800 800 0 0 0000 0 0 0 0000 DNAT O OO O O D FUJIFILM 0 0O O 800 0000 00000 00 0 0 000 0 0 00 0 0 000 0 00 000000000060 0000000 0 639 07151 QuickGene 0 O0 0 50 960 0 25550001 13 250 00000 0 COL 5 5 1 10 0000 00000 D 1
16. 00000 00000 0000 00000 00 0 0 0 00 00 L OOO0OOOCOCOCOOCOOOOCO OODOD00HK 00000000A 000 00 00000 00 0 0 0000 000 0 0 0 0 0 0 0 00 26 47 0 00000 00000 0 0 0 0000 00 00000 0 0 0 0000 000 2 2 10 0000 ec 12345678910111721311151617181920212223245 2722930313 f 2 6 4 7 000 10 00000000000 0 2100000000000 0 3210 0000 0000 High Throughput Westem Blot
17. OCOL 0000 gt lt lt lt gt lt 5 69 6 gt S lt 02 620 O 00 0 0 00000 O L 0 000 000 0 0 0 0 0 0 00 0 0 0 0 0 0 00 0 00 0 0 00 1 31 55 1 0 0 0 HDAC Cla s 2 0 00 00000 HDAC 1 p0000000000m0000000010 4 7970 0 0 0 0000000000 9 000 HDAC 0 0 M344 03 307 39 292 12 N 1293 28431 MS 275 37641 N H 2 NN n OH 2 CH s Piceatanndi 0 244 24 2 394 47 HO OH HO T NJ OH NH 198 22 CH3 00 0 0 0000 0 10000000
18. 201 1 20091 0 000 40 00000000 98 0 8u i N L 210 00000000 99w w 0000 12 10000460300 0 2 0 0 0 0 0 000 2100000000000 20 0000000000 A Bio 0 0 O0 DMM 0000000000 Wako Bio Window 0 00000000 200000001 Wako Bio Window 0 0 0000000000000000 0 0 0000 000000 000000 Anaytical Circe Organic Square 00 371 91 91 00000 0000000000 00000 E mail biowin wako 06 6201 5964 14 Wako BioWindow I AUG 2005 No 69 1 Transforming Growth 1 Human Insect Cells recombinant 0 Human transforming growth factor B 1 cDNA expressed i BTI Tn 0 00 ED50 lt 0 05 ng m 0 0 0 2x 107 0 5B1 4 High 5 insect cells 2000 00 0 0 0 0 0 It 4 0 6 000 0 0 0 0 00 1 207 16241 00000 00 8100000000000 _________ g 3900 0000 0 00 204 14291 0 0 00000 0 000cHog 0 0 00 0 49 000 K T 0000000 m O Wako Connective Tissue Growth Factor Human recombinant 00 Human
19. _ 9000 207 13463 recombinantMouseTNFa ____ 550000 Wako Bio Window I AUG 2005 No 69 000000 000 000000 20000 0000 000 2 10 e0001m 51000 0000 00 0 00000000 90000 8 120 0000000000 500 aore 0 0000000000000 000 j 2 PIME 8000 0 1 sO IO 100m ob ob Mouse Insulin Resistn 0 goong Insulin 6 84 L 610 92 97 i 2 42 3 42 71 3 91 8 000 0 77 96 458 97 2 57 3 72 ng LF Resistin ng m ob 1 0 0000000 0 0000000000000 100000000 00000 0000000 0000000 000000000 0000000000 000 O KK Mouse
20. 0 00 A 0 0000 Sample Resistin ng m Sample Resistin ng m KKJO0 6 455 KKJ 00 1 4 30 OD 7 8 78 00 12 6 59 8 9 44 14w KKJ00 B 3 46 kK 00 9 9 06 ____ ___ 467 _ 10 6 68 mean 4 76 mean 7 70 SD 13 SD 21 000 0000000 ________ ODO 00000000 63207141 AKRRS 011 000 00000 000 62 000 00000 ______ _______ 00000000 000 00000 000 48 000 633 03411 031 000 00000 000 4200 Wako Bio Window AUG 2005 No 69 00000 00 Miser Antibody Extender Solution NC DO0000000000000000000 gopoo0o00o0o000 0000000000000 10 12 000000000000 0 10 V12 LIUS 9 219 39 419 10 000 000 0000 00000 558 71805 32110 Miser Antibody Extender Solution 8x 100 0207 500m 556 71801 32105 Miser Antibody Extender Solution N Trial NCI 8 10002 0 50m 0 0000000 Qentix Westem Blot Signal Enhance 210501 0 00000000000000 0000D L Biotechnology Caspase Anti Caspase 3x Glutathione S Anti GST NFx B Anti NFx B GxHistidine GFP Ant
21. 0 50001 100 0000000000000000 0 5 L Testeri ONA 0 0 00000 000 0000 00000 0 0 0 0 000 00000 0 0 O 1 00000 Sense 0 0 0 0 Antisense 0000000 0 000000000000 20 0 0000 00 00000 000 DSN Duplex specific nuclease 00 0000 2120 0 00 00 00 00 00 0 0 000 0 00000000000 Primeri 0 5 1 00000 000 1 Tester 00 0 400 00000 0 0 00 0 0 0 0 0000 E 2 02 0000000000000000 0 00 Tester 55 Driver 00000 000 00000 00 0 0 0 0 0 Dives 2001 0000000000000
22. 0 00170110 0 100 000 170 40 00106060 3 6 TELO 03 3219 2211 00106060 00 00 0000 OOOO 0 0090000000010 00 00 1 O 0000 0 0 000 10 000 0000 000 gt gt 10 050 00 00 10 100 000 10 500 000000000000000 0000000 00 On 11 300 000000000 OS 12 100 00 13 05 00 00 13 450 00000000000 0000 00 00 14 250 DO00000000000000000 0 000 00 00 15 050 0000000000 15 200 00 00 16 000 50 000000000 0 000 16 400 00 00 17 200 0000 00 00 17 300 0000 000 0 0 0000 0 0 4200 00000 000000 000 0 0000000 103 0023 00000000000000 50 130 03 3270 8582 03 3270 8243 E mail seminar wako chem co jp 46 66 46 6060006600006 46 646 646 6006606600600 600600600 6006 600606 000 600600600 600600600 000 0 469 2 226 26 26 26 226 26 26 626 626 626 2 226 26 26 626 626 2 2 226 26 626 626 26 2 226 26 26 626 626 26 2 2 2 2 26 100000 000 0 0 0000 000
23. 000 0 0 000000 0000000000 0 000000000 0000 Saccharomyces cerevisiae Homozygous diploid deletion 565 Invitrogen 2540LH1R3 84743 background Plasmid YEp51 LEU2 2u Selection platg L leucine drop out plate No 0301 51 LO No 0302 51 L 0303 51 i _ 000 0 000 00 0 0 L 0 00 00 0000 0 00 0 gt 00 000 000000 00000000 00000 UG OO 2005 100000060 5 480 0 1 0 00 000 0000 K O O DNA Step Ladder
24. 637 05631 37 400 638 07001 RT S QuickGeneRNAD 0005 __ 0 5140 639 07151 PL S 00005 _ 0 B5 2508 0 00 Wako Bio Window AUG 2005 No 69 G K com Anti Mouse 196 000000000 0000 0000 e0 O 0O 0O00 0O 000000000 000 Cinni 0 0 0 0 m 0000000000 manara o000000000000 m 000 0000000 6410511 3471 00000000000000 0 0000 48 960 647 0517 373 00000 0 000000000 0 0000 49 920 644 05181 3474 0000 00 000000000 0 0000 _ 5240 641 07391 3261 000000000010000 2 2800 644 07401 3262 00000 0 0000000000 200 30000 000000 00000 0000
25. E E E E E A E p 6 5 5 O Genoglass PGCOODO O 7 High Throughput Western 32100 D p 8 BioChain Frozen Tissue Array 9 246 20 000 Break 14 OOL 20050000 7 poopoo 0 11 pti 00000000000000000 000 rees p 12 00000000000 siRNA 19 p 23 210 000000000 p 24 24 O DsDD cDNA Subtraction Wako DsDD Duplex specific Direct Digestion Subtraction Kit O O O 0 0 0 cDNA Library 0000 Tester 0 0 Driver 0000000000 0000 Tester Driver 000000 DNAD 0 0000 0 00 Duplex specific nuclease 0 000000000 0O 0O O 0 00000 00 Driver 0 0 00000 0 0000000000
26. 0 139 14671 M34 0 0 DTrichostin 00000 mg 19000 093 05251 ITSAL Trichostatin AJ 0 0 0 smg 000000 7 500 136 14681 MC 1293 0000000000000000 smg O00000 18 000 132 14661 5 275 310 00 8 0000 13 000 169 21661 Piceatannol Classlll HDAC 0 0 0 0 000 0000 9000 18 000 197 13671 HDACO 0 0 0000 0 0 00000 25 000 190 13661 Splitomicin 0 0 00000000000 7 000 K W Wako Bio Window I AUG 2005 No 69 13 2 20904 00000000000 000 x 4000 00 ww 7 100 0 000 2 oO000000000 114 00000000000 0 000 00000 7813 1114x 1 000 mpx 100 w w 100 78 13 14 26 000 10000700 00024 moi O O OO NI 00000
27. 0 0 000 00 0 06 0 000000000 00000000 0000060 00000 a 00000 0000 000000000 0 iPod 00000000000 00000 00 00 0000 6000 100000000 0 9 00 0000 000000 00000 000000 00 0909 000000 008 00 0000 0 00 0 000 0 00 0000 00 000 Wako Bio Wako Bio 000 0 00000 0 E biowin weko F A X 06 6201 5964 G K Oo U L a 9 141 16 DD 10 50 7 10 190 22 12 0000 0000 0000 00 000000 0 2 4 0 0 0 2 41 0 0 0 0 158 0 120 O Argi 00000 2 4 2 0O 000000000000
28. 0 0 00000000000 lt Alia 4 76 000 000 556 6551 0 0 0000000000 0000000 586 97821 _____________ 552 65891 550 65951 559 62981 2 gt VD 005 Recombinant Human VEGF D ____ 555 62961 321 1 050 Recombinant Human VEGF RY Y Fc Chimera __ 552 62971 357 KD 050 Recombinant Human VEGF 82 KDR Fc Chimera ___ 557 72291 349 F4 050 Recombinant Human VEGF R3 Flt 4 Fc Chimera ___ 551 65501 293 Human VEGF Affinity Purified Polyclonal Ab ____ 1009 550 65711 752 Human VEGF C Affinity Purified Polyclonal Ab ___ 104g 557 75591 286 Human VEGF D Affinity Purified Polyclonal Ab 553 65681 AF321 Human VEGF RY Fit 1 Affinity Purified Polyclonal Ab 1009 550 65691 _AF357 Human VEGF 2 KDR Affinity Purified Polyclonal Ab 1009 552 60151 AF349 Human VEGF R3 Fit 4 Affinity Purified Polyclonal Ab 1009 554 6011 550 75601 550 67151 558 60131 555 60141 MAB3572 Human VEGF 22 KDR MAb Clone 89106 ____ 5g 558 73941 551 60121 _ __ Human VEGF ____9_ _ kt 559 60161 _ Human VEGF D Quantikine ELISA Kit ___9__ _ kt 586 99881 100 Human RY Fit 1 Quantikine ELISA Kit __ lkt 55173311 DVR200 Human R KDR Quantikine ELISA Kit _
29. 0 68 000 4 IE 35x 108 0 00000 000 200000000000 D00000000000000000000000010 gt 0000000000 0 S DO000C0O00000000000000000000 ira DO00000000000000000000000 0000 0 000000 00000 000000000 0 0 0200 0 00 0 0000000 3 5 10 00 2550mg 0000 0190 0 0 100 20000000 000 DELSA 000000000000 0 0 0 OSELDI 000 0000000 ________ o O 551 71611 444810 0 00 000 0000 61 900 000 000 0000000 a a 556 71541 122640 0 00000000 31 000 553 71551 36000 550 71561 49700 557 71571 53 300 554 71581 55 700 551 71591 55 700 554 71601 58000 558 71621 61400 555 71631 63700 552 71641 63700 559 71651 45 600 556 71661 44 900 0000D L Novagen 100 000 000000000
30. 00 0 00 0 00 00 0 0000 00000 00000 0000 000 0 00000 00 0 00 siRNA 173 000 gt 10 200 41 OFF 173000 102 000 OFF 1 41000 I S Wako Bio Window AUG 2005 No 69 20 0 00000000000 0246 O Neuropeptide NPY 0 000000 Neuropeptide 00000000000 000000000 n vitro 0 0 0 00000 0000 0 ICs Y Ys OOO 10 000 15 10 000 10 000 2268231118 1 000 3 3 1 000 1 000 1 al 384 1999 21 al 129 1075 2000 O n vivod O O O U DO 0000O00000000 Y0000000000000000000 000000000000 000 0000000
31. 00 00 0 00 0 0 6 6 Exonucleasel 11 0 000000000 0000 00 000 000000 20000000 Subtracted cDNA OUUU 10000 0 5u x 11 s 10x Lambda Buffer 25u 1 s4x Hybridization Buffer 25u x 1 s Duplex specific nuclease ssssssnnnnnnnnnnnnnnnnnnnnnnnnnnn 5u x 1 s 2x Duplex specific nuclease Buffer 25u x 1 5u x 1 s10x Exonucleasel Buffer 25u x 1 nin 50u 1 25100 50u 11 s3mol Sodium Acetate pH 5 2 200 1 SOARE arsana ENA 1 Q 500000000 5 Duplex specific Direct Digestion 2 0000000000000 Duplex specific DNA 1 Testeri 000000000 0 2 11 Driveri 0000000000 000000 Real time PCR
32. 00 8 1 100 9 Helal O 00 000000 Catch amp Release Non Denaturing Elution 0 0O t 4 15458 000000 Catch amp Releasef Denaturing Elution Buffer 0 0 0 00000 00 0 0000 0 00000 00000 000 00000 E 45 10 0000000 00000000 00000 L 000 22 5 620 00000000 20002 00 TAH WO OOOO O O00 00000 E Tg 00 6000 70 U 7 5 00 000 OFF 000 988 00000 0000000 00000 00 OFF 00 OFF OFF 00 OFF 00 siRNA 2000000 0 00 0000 0000 00000000 515 0 000000000000000 0 0 0 000 0 0 000 00 0 0 00 0 0 0 00 00 00 Smith Waermani 0 6 50 1 0 000 0 0 0 0000 0 000 0000 000 000 0 0 0 0 0 00000 0000000 00 0 000
33. D et al 1997 J Biol Chem 272 25176 00 0 0 133pg m 2 Achen al 1998 Proc Natl Acad Sci USA 95 548 0 109 7000 pg m 3 Lee J et al 1996 Proc Natl Acad Sci USA 93 1988 4 Siegfried e al 2003 J Clin Invest 111 1723 5 McColl B K et al 2008 J 198 863 sVEGF C 96 80000000 10 EMBO 16 88 5 0 HRP 21 is N E al J Biol Chem E print T 70 Kukk E et al 1996 Development 122 3829 SE 11 10 Makinen T al 2001 EMBO J 20 4762 5 21 u Saharinen etal 2004 Trends Immunol 25 387 2 12 Nisato R E al 2003 Thromb Haemost 90 591 aa 6m 13 Stacker S A et al 2002 Nat Rev Canc 2 573 000 0000000 ________ 0 o 554 74261 Human VEGF C Quantikine ELISA 00 0000000000000000000 0 0 50 6570
34. ____ Ikt 556 65051 5008 Goat HRP DAB Cell amp Tissue Staining Kit 5000 00 0000000000000000000 http mdsystems com 58 000 58 000 60 000 58 000 73 000 62 000 73 000 73 000 73 000 73 000 73 000 73 000 73 000 60 000 60 000 60 000 60 000 60 000 60 000 76 000 76 000 76 000 76 000 40 000 U TN ProteoExtract KitD 0 0 0 Calbiochem mO 1 10 OUL gt 0 000 9 0 gt 0000000 g k a E 7477 BRAHEA 1 paris EHE 3 i 5 6 E __ 00 0 000 0 A4310 0000000000 15 0 000000 00000 0 p 00 00000000000000000 000 000000 _____ 00000000 579 77231 539790 0 0 0 000
35. cDNA expressed 0 g O in E coli 00 EDS0 1 0 2 04 0000 0 000l 0000 11 200 981 000000 000 No Ooo 0000000 036 19471 CTGADODO 000 39 000 K T 00010 O00No 00000000 033 19481 CTGFL 00 0 000 39 000 O OUO UL 00000 __ 00000000 CDP 098 04623 recombinantHunen IL 3 6000 090 08963 recombinantHumanIL 4 7000 CD 096 04283 recombinantHunn iL 9000 097 05173 recombinantHumen IL 13 90000 CHD 067 04053 9000 CD 137 13614 0000 188 01473 recombinantHuman sRANKL __ __ mw 199 12813 recombinantHuman SCF 201 15264 recombinantHumen TNFa _______ 30000 CED 229 01313 recombinantHuman VEGF _ _ _ 000 41131 090 04683 recombinant m 9000 096 08943 ______ mw 7000 094 05183 _ ____ 90000 CHD 079 04673 mw 85000 CD 13114393
36. i GFP 4x Ubiquitin Ant Ubiquitin 10x 00 p53 Anti p53 1 Bx 20 IL 6 Anti IL 6 25x Cyclin E Anti Cyclin E 100x Phosphotyrosine Anti PY 20 100x pi Antibody h0 Extender eko 00000000 000 5100000 10 0 50 0 0 000 000 430 0000 0 0 0000 1 2 3 4 5 1 2 3 4 5 1 2 3 4 ez gip Oo H __ __ 1 1 2 amp 4 amp 2 0000000 0 000000000 0000 21 900 8 100 000 0000000 00000000 OUUU 531 50251 34075 100m 7 ar E 53484551 34095 o 100m SPESI Fpa 00001 527 79775 21059 Restore Westem Blot Stripping Buffer ____________ 500m 67 900 108 100 68 800 113 200 15 000 44 900 Wako Bio Window AUG 2005 No 69 0000000 ea upstate Catch and Release v2 0 cell signaling solution Reversible Immunoprecipitation System Upstate 1 EE E EE E E
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