112号 pdf版
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1. artificial light 611 ole NOD 001 4 nne vr natural light intensity Ix 10 108 10 10 107 109 10 104 106 power W mm 10 10716 1074 10712 10 10 1078 1079 10 102 Fs 10 10 104 10 108 101 10 1014 10 555 photon mm s Fig 3 Schematic diagram of relationship between light source and photonic level 22 OOOO 10 lamp light am 100000000000 plification by sim2. a 5 2 Mm dE c E gt CO o CC 2 rr o 3 L3 g E 2 c E 5 m r9 6 Ea r3 5 BE de E ON 3 8 E 5 9 a ES 0 g Es r3 Ei r3 num r3 a 2 E x oH 2253 s E Y m es E 9 r3 r3 3 pg 2 r3 0857202 ogogpgs 3255 CH e 5 5 E E La P3 us 68 Am JE J 1 E H000 gt EES r3 amc 3 o r3 CH oo E 5 es s ua EE S 22 5 ea 1 SE E SE ca 3 Or 3 3 22 5 Depp 7 o 9 2 L1 6 a c 8 2 gt S Ee
3. 4 O l J OTsukatanig D 0 0 0 2 3 5 6 0 00000 1 4 000 4 1 p00000000 0000 1 pH 7 00 pHo 980000000 wst1g 111 000 0000 000000000000 OOO OOOOOOOO 1 1 11100 0 0 ESR0O000000000 0 0000 020000 wst1p0o000000000000 000 WST 1ig 000000000020 0000000000 pogo ooo Wst14go00000000000000 1111111110 1 ago o 000 000000000000 DO000000000000 20000 0000000000 000000000 000WST 100000000001 0x 105 DO00000000000000000 gg 2 0 100 DO OC 000 000000 90010 0 0000 ego 880000000020 000000000090 1110 pop 0000 0000000000000 020 0 0000 000000000000
4. pogo ooo 000000000000 00000000 SOD0000000000000000000 00000 Fig 920000000000 WST 10 wsrigppoo 0000000000 00000000000 000000 gt 0 0000000000000 000 ODO ICso0 0 0 SODOOOOOOOPFOOOOOOOOOOO 58 10 0 000 000 0000 11 000 0000000000 5 10 00000000 5 3 O 9000000000 1 SOD00000000000000000000 5000 02 110 0 0 00 0 0 0 00 0000 111111 111 DO000000000 0 00 000000000000000 DO000 000000000000 0 SOD Assay Kit WSTO O DODO 0000 000000000000000 020000 111111 1111 111 11111 110 0 SOD I 105 OO 2 0 5 0 00 00 D000 2 010 000000000 0000000000000 DO000000000000000 50 000 ESRO D 20 1110 08 10000000 0 0 50 000 11111 11 1 11 DO00000000000000000
5. 000000000000 000000000000 1 M0 000000000 00000 shift enpnn nna anggagddtgena npna npnann 1111111 111 111 1111111 111111 1111 111 spatial resolution O O 0000000 Fig 0 00 0000 optical microscoper 0 111 11 0 0 0 0 0 400nmp 000000000 111111 1111 111 1 111111 1111 111 0 O contrast enhanced microscopy 000000000 000 phase contrast microscope 00000000 differential interference contrast microscope 000000 OOOOOOOOOOOOOOOOOOOAO 400 1 1 000000
6. 080000000 Fig 300 0 00000 0 00 10000000000000000 pwst 190 0000 0 0wsT 1 ca 000 JUD pego 0000000000000 02000000 pon 000 0000000000000 WST 10 10 WST 10000 0 0 438 0 000 D JO 020 1100000 0 458 1000 Fig 400000 000000 FIAD Ton 850 nu 100000 Sepharose XO 000000000 5 1 O 000000000000 01010107600010 1000000 0 111 00 00 00000008 00000 1 1 UD DO00000000000000000000000 WST 1 poo 9010000 0000 D000 00000800 000090 000 1000 11 0400000000000 WST 1000000 0000000 0000 1000 6 0 DO 0000000000000 WST D00 6100 1000 Uu D000 100 10000 0 10 20 0 4 ml min 0 0 0 0000000090 0003004 mMo0000000000254Mg 1 0mMgg 0 0 0 0 D 1000 NOOO 0 O O Table im 11 1 1 00008 4 3 02 0000000000000 0000 1000 O BD 0 5 mM n 1000 11111 00 0xopo 0000000000 100 010000030000 000000
7. 0000000000000000000WST 19 SOD 0 000 100000 10000000000000000000000000000 0000000000000000ATPg 000000000 00000000000000 00000000000000000000000000000 0000000000000000000000000000 100000000000001000 500000000000 000000 100000000 000000000000 70 90 0000000000000 0 0000000 0 0000000000000000000000 81 0 0 0000000000000000000000000000 D00000000000000000000000000000 10000000000000000000000000000 00 00 0000000 000 0000000 0000000000000000000000 000000 D00000000005000000000000m0000 200000 000000000000 000000000000000 000000000000000000 00000 1000000000 400000000000 0000000 0000000000000000 000000000000 00000000000000000000000000000 0000000000000 0000000000000000 000000000 10 000000000000000 e e e e 000000 0000000000000000000000 O22 Oz gt H202 gt OHD gt H20 DO0000000000000000000000000000 10 000000000000000000000000 00 000000000000 co000000000000000 1 0 00 55 8 0 01101 600000 00000000 080 200 50 00 0 000 0090800000000 0000000 000
8. 75 100 WST 10 D 100000000000 SosAnn 1110 400000000000 1000000 5035 0000 Table 30 10 1 ESRO 0 0 SOSAg 0 00 WST 10000000000 10 1 000 0 0 958000000000 5 50 000000000000000000000000000 5 JU WST 10 0 ESR 0000 t 0000000000020 U uuu JU oo 10000 EIU U 10000 25 4 3 2 log 1 dilution ratio O OSOD Assay Kit WST Fig 10 Inhibition curves of some food samples obtained by the WST 1 0 10 0000000 10 10 10 method The food samples were green tea O red wine rJ 20 5 35 n coffee A and cocoa 0 O SOSAJ O ut 00000 1 110 00 Ka Table 2 Comparison of SOSA of some foods measured by WST 000 5 1 1 and ESR method 505 0 5 00 000000 Method sample ESR WST 1 Red wine 1 units ml 866 348 61 uU U Red wine 2 units ml 832 392 WST 100 00 0200 11101 0 0 Red wine 3 units ml 755 199 5 11 0 000000000020 Green tea 1 units g 67 100
9. EO MUD 0010101 Bg 9 45 10 30 TU o UO TE O DU o aa 0000000000000 000000 00000000000 200000000 ga a 10 30 11 15 o 00000000000 0000000 11 11 po0000000 ga ga 11 15 12 00 pad 00000000 0000000000000 TUD adn da 2 000000 11 13 15 14 00 a 14 00 14 45 Ua 1111 1 1 1 ARO ORD 200000000000 pa ga 14 45 15 30 pana o 0000000000 MUD DET O 3 00000 ga ga 15 45 16 30 DUD 0000000000000 ga ganada 11 11 11 1 TD 1 1 000000000000 16 30 17 15 o o T DOU TE U o o o U 30 2025 5 0 Tel 0120 489548 Fax 0120 021557 E mailiinfo dojindo co jp ennmnn nnnnnnnnnnununnnnnmnnntim 17 45 19 30 O
10. 00000 0000 ONOD O 0000 Bacillus licheniformis 0 0 0 0 0 8 1000000000 00 11 0 00 000 0 0 0 0 Tirong OOOO U DO000000000000000 00000000000 0000000000 0 O 9 00 NADH 000000 0000000 0 50 50 4MO J 0 0NADHO O 438 nmp 11011 Fig 1 1 000008 DODO 0 0000000000 1 Fig 7200 0000000000000000 0 000 0 0670 168 yMODO000000000000000000000000 1 1 0 0 000000000000 0 00000000000 00 100000000000000 00000000000 080 0 0 NAD X X Acetaldehyde NADH O2 WST 1 formazan ADH NOD Fig 7 Reaction mechanism of ethanol analysis
11. pog8080000000000000000006FPg00 In vivo labeling 0 O0 0 0000000000000000 280000 000000000 000000000000 00000000000 0000000000 8 in 0 000000000 O in vivo 90 001 1 000000000000000000 000000 000 000000000000 65 000000000 DAFRO 00000000 PAlaPAla 00000000 00 Ka 0 1000000000000000000000 1 1 0000000000000000000000000000 0000000000000 Dio obogonalt O OOO OUD Tsienn 200000 FAsHo oo 0 00 0000000000 000000000000 ccxxcca xo 000000000 1110 0 0000000000000000000000 200 111 0 0 0 0 5 0000000000 0 E lt lt BGFL Fig 2 hAcTO000000000000 14 DOJI News No 11
12. 1111000000 0 0 00 54 9000000090 1 00000000000 000000000000 emitted 0000000000 01 000000000000 00 excitation light D O 0000090000000 00 Jablonski diagrama OOOO stokes 6 10 00 100 6 O fluorescence life time OD 10000 0090 epo000000000 gt 5000000000 quantum yield 000000 000000000000 6 tetramethyirhodamineon 0 0000 000000000000 00000000 evanescent wave 00080000000 1110100000 0 0 0 spol 000090 000 Fig 000000000000000000000 gt OOOOOOOOOOOOOOOOOOOO multi photon excitation O 0 0000000000000 8 1 1 0 0 00 0 UU fluoresceing D OOO 10 00 1 1 0 00000000000000000000000 p0000000000000000 fluorescence resonance energy transfer FRETO 0000000000000 000 0 1099000
13. 6 03 1 E s Bi oooo OJO 5 MES V Ej a S s E ES ra ng Del EM CC pugpoHaugnugd BSS lt 2000 S MET E CC CH 2225 2200 1392 2 z og ooo gt ou CU Do o E E 3 3 ooo L3 d Dro c E T CO r3 20mm 35 oa 2 7 gt oo 9 oft a 3 Pon O Z 8 8 2 S Lo ooo Poo O 5 5 o 6 oooo o CH ra oh c NN E Ska GSS oO D cg e z mn 5 SI 30 12 222 23 A r3 SL e 5 5 CC Ou HN 6 Vez 8 g a m N gt P Cool 2 5 DN 5 r3 o lt 5 n gt Do 2 3 E u 65 NPOODSTODO y 2 2 E 8 4 CDS 9
14. 0000000000000 Fig 2000000000000 pggd8 0000000000000 000000000000 pg 280000000000 000000000 000000 DODO 0000000000000 000000000000 0000000000000 000000000000 gon 0000000000000 0000000000000 32 00000900000 DODOM 1 1 10 110 0 0000000000000 Young 8 Holmheltz 18520 1 1 11 1 0000000000000 11 11 1
15. 0200 4 2 00000000000 02000 DO00000000000000000000 U O Chattonella 0 00 00000 0 0000000 00000 0 antiquajj Chattonella marina Heterosigma akashiwoy 000000000000 Skeletonema costatum Gymnodinium mikimotoip 000 00000000000000000000000000000 5 11 O po0000000000000000000 Po 000000000000 DO0000000000000000 000000000 111111 11110 111 wstr19p0000000000000000000000 doo 000000000 000000000000 020000 DO000000000000000000 5 1110 0 0000 pgp00800000000000 Chattonella 000 H 00000000000000 0200000000 WST 10 akashiwo 0000000000WstT10000000000 11 0 000000000 0000000 37 C 438 1 1 1 0 00 1 200000000000000000 0 4 2 mM WST 1 SOD0D0000000000000000000000000 0000000000 10m0 000010000 458 0 poa 30000000000 O 000000000000 OOOOOOOOOOOOOOOOOOOOOOAO D00000
16. 0 000 111 0 8 8 0 0 00 000 00 1 11 0 Tirong 9000000860 t00p 0000000000000 1111111 1111 111 DO0000000000000 020 000000000000 1 1 9 10 000000 000 Du 000 6 0 SODO Tirong 00210 4400000 0 1 0 0 3 E E 5 B 0 2 o 0 05 _ 8 8 01 5 2 lt lt oc e e s s 7 0c Incubation time min Incubation time min Fig 2 Absorbance change by WST 1 assay Glyceraldehyde 5 mM N Fig 3 Absorbance change by cytochrome cassay Glyceraldehyde 5 mM acetyl L lysine 10 mM and DTPA 50 uM in 50 mM chelex treated N acetyl L lysine 10 mM and DTPA 50 uM in 50 mM chelex phosphate buffer pH 7 0 were incubated for 2 days at 37 C The treated phosphate buffer pH 7 0 were incubated for 2 days at 37 assay was performed in the absence O and presence of C The assay was performed in the absence O and presence SOD e of SOD DOJM News No 112 2004 Table 1 Superoxide Anion Generated by Various Carbohydrates with and without 10 mM N acetyl L lysine N Ac Lys Superoxide anion formation nmole ml h Carbo hydrate WST 1 method Cytochrome c method 5 0 mM N Ac Lys Aldehyde alone
17. OO gamma ray 11 1 0 0 0 E EJ ELE ELE E E E C Et O a LJ 3 LJ CJ 5 LJ LJ o EJ o A L3 r3 C3 C3 C3 p 000000000 LJ O infrared IROD 1 UU visib l OO OO E o 1 EM length m LB 10 radio TV 10 86 micro 104 tissue cell 10 6 visivle orgenelle UV molecule 8 10 DNA 10 atom 10 12 10 14 10 16 Fig 1 Scales of electromagnetic wave EM and living bodies LB arrows a theoritical limit of spatial resolution by a human eye 10 mm b optical microscopy 107mm c scanning probe microscopy or atomic force microscopy 10 mm and e transmission electron microscopy 10 mm UV Visible IR violet blue green yellow orange red gt Relative sensitivity o al 555 400 500 600 700 Wavelength nm Fig 2 Wavelength of visible light and sensitivity of human eye o DOJIN News No 112 2004 photon counting fluorescence transillumination cellular images
18. 02000 onga sos 63 63 63 3 5 pj 53 C 9 3 2 p OR 2 Oo 502052400217 lt am 3 2 252 3 22 as Eu c E C 550255008 SN E E aer SE ler Een EE 0 EE Gagan 58 5 33 Eat KE EE 32 3 3020595590005 25050550500 D y DO ET ES Tis 2222 222 222 nuugumnguurn EE 52 6363 55005 3 3 3 03 32 A 020 o DD 32222253 222239 2222 2 2 6 6 22 2262 2 2 909 22 2 56 220059422922 9222205
19. 220004 AA 52 53 2 cuntur Ss t E EEN 51 00 61 1 0 E EL 51013 51 SO ricguuuguH muuuut 5205022000500 000000000544 0000000 025252008522 r3 r3 ET c EX EY ES 032 BDSG EE 5 e EEES Door 55 Oo OS 2 DODODOS 3 326 3 2 o x CCS EJ Poong B Lir a a EXIT E3 53 jlo ES 2 E O E e EO E E DOJ N News No 112 2004
20. 0 0808 00000000000000000000000000000 sob OO 1 000000000720 0000 10 00000000000000 100000 poo 00 08 11111 1 DO00000000000000 0 0Mn SOD00000000 11111 1 1 2000000 100000000 3000000 1111 00 0000 00 0 908 0 00000000 000000 3000000000 SOD Assay Kit WST 0 Mn sOoD CC 0 00 Mn SOD 0000000 SOSAD 000 00 Fig 190000000 500 0 1 1110 0 0 000000 0000000000000 1111010111 0 0 1800 2300 00 0 0 1110000000 Mn SOD00000000000 000000 2800000000000000 210000000 WsT i0Q000000000000000000000000 DO0000000000000000 00 WSt 10000000 ag a ooo 000000000000 11 1 1 1 280000 00 1001 000 0 500 000000000000 0000000000 00 0 80 0000000 Germing 0 0 0 0 0 000000 00 00 0 0 0 0000 00000000 SoSAg Table 2 n a ESRB 0000 000000000070 000000000000 ugpnagnawsr343pgagg 0008Woo0 000 Ger
21. CONTENTS o Review wWwST 10000000000000000000000006 LB DS A eno ne 11117 G Topics on Chemistry 4 000000 In Vivog ue 4 00000 2004 ISSN0385 1516 DOJM News No 112 2004 DD Review wst1p00 0000000000000 000000 111 ABBA 1 000000000000000 2 11 111 1111 100 1010 0101 1 OOOO 9 Topics on Chemistry 000000 Am BAA DUG crs 14 Commercial 00000 16 s l ites on nies 19 ee 20 EEE HEEE ee a 21 Q amp A 104000000000 HEH 1 ss suu aaa 18 una 13 11 1 1 1 UUO utin 13 TR RR ECH ua usu RE 15 22 po00 1001 000 0 SOD Assay Kit WST sass 8 Nucleostain DNA Damage Quantification Kit AP Site Counting 15 Alkaline Phosphatase Labeling Kit NH2 16 Alkaline Phosphatase Labeling Kit SH 17 Biotin Labeling Kit NH2 4 9 99 9 9 99 19 Biotinylation Kit Sulfo OSu 19 gt DOJ N News No 112 2004
22. TELFAX E mail 0 00000 E mail FAX 0 00 0 009 0120 021557 URL http www dojindo co jp E mail info dojindo co jp EASES 00000000 No t12 00160 1001000 000000 DOJINDO LABORATORIES poyin News 2 P O00000 080 8088 Oooo 00000 0000000000
23. WsT 1000000000000000000000000 Detection of Superoxide Anion with WST 1 and its Application Summary Superoxide anion O2 is formed by one electron reduction of molecular oxygen It is a precursor of highly toxic hydroxyl radi cal and has a high reactivity with nitric oxide to form peroxynitrite The detection of unstable Oz is generally based on indirect oo UU method using a probe which can trap O gt to result in any chemi O Hiroyuki Ukeda cal change There are several kinds of probes for Oz such as 000100 colorimetric spin trapping fluorimetric and chemiluminometric probe Among them colorimetric probe is the most convenient one because of no need for special instrument Instead of its con venience the conventional colorimetric probes such as cyto chrome c and NBT has a drawback in the selectivity In order to overcome the problem we examined some water soluble tetra zolium salts which have been recently synthesized and found that WST 1 shows a satisfactory property as colorimetric probe be cause of the reactivity with Oz and the selectivity In the present article describe the reaction mechanism of WST 1 with O gt and application of WST 1 to detect Oz derived from biochemical materials such as animal homogenates microalgae and yeast and to estimate the scavenging activity of O gt which the enzyme superoxide dismutase and some bioresources including food components have
24. A12 D000000 mgo 0000000000000 Peroxidase Alkaline Phosphatase 1 mg 10 mg es pagada na OOOOOOOOOOOOOOOOO A5 UU Storage Buffer 0000000000 20000 Du 00000000 Tel 0120 489548 Qe Ol O 6 000 g 190mingaoo 00000000000 Fax 0120 021557 000000000000 E mail info dojindo co jp A6 00000 111111 8 DOJAD News No 112 2004 8 1111111111111 Biotin Labeling Kit NH 0000 20000000000000000000 e NH gt Reactive Bioting 0 00 e Filtration 06101 01 00000 010001 1 0 30000000 OOOO IgG U0 NH2 Reactive Biotin 1 IgG NH2 Reactive Biotin 3 Reaction Butter 0 5 ml x 1 rod WS Buffer 4 ml xip Er HUI Filtration Tube 30 one Biotin Labeling Kit NH2J 00 00000000000000 0000000006 1 0000000000000000000000000 NH a Reactive Biotin 00 0000000000000000000 00000000000000000000000000000 0 0 0000000000000 9600000000000 J D 00 D O Filtration Tubeg D D 00 000000000090 100000000 0000000000000000 0 000000000
25. 00000000 0Q00000000000000SH000000000000 000000 e Filtration Tube D 0000000000000000000 1 00000 AP0000000000000 S S00000 0000000000000000000000000 ao NA 008 0000000 30000000 SH Reactive ALP 100 ug x 30 00000000000 Reducing Agent 80 ug x 30 Solution A 4mlx 10 Solution B 1mlx 10 aii ALP Reaction Buffer 200 ul x 11 Storage Buffer 4mlx 11 Filtration Tube 3 pa D IgG 3701000000 ES 0000 Alkaline Phosphatase Labeling Kit SH SH O O O U U ug SH Reactive ALP 0 uggagnagaudg 11 0 0000000000000 0 0000000SH 110 0000000000000 SHOO 5 ALPO IgG 11 0 0 00 00 0 00 SH 11 0 00 0000 0000000 00 95600000000000000000000000000 Filtration Tube 00008000 Fig 20 0000000000 Filtration Tube 00000000000 1 10 1 11 10 1 000000000000 Filtration Tube 4000
26. UD welg 000000 00 6 HRP streptavidinQ 000000 PBSTO 1 50000 U 00000 O O HRP streptavidin 000000 PBSTO 1 4 0000 gt lU UU UUUMUOU DODODOO Nucleostain DNA Damage Quantification Kit AP Site Counting 5 samples 22 000 DK02 20 samples 69 000 DK02 News No 112 2004 96 111 E 1111 E 11111111 A Alkaline Phosphatase ALP O 0000000000000 D0000000000000000000000ALPO DO Peroxidase 0200000000 Enzyme Linked 0000000000000000000 Immunosorbent assay ELISA 0000000 DOU ALP 00000000000000000000000000000 000000000000000000000000000 r 0000000000000000000000000 NS Y Alkaline Phosphatase Labeling Kits O O O O O 000 90 yn HO0000000 ALPO000000000000000 E ua l Reactive 0 00 O sos NH2 Reactive ALP Bro pg00000000000000000 G 96 CE NH2 Reactive ALP LP LP 0000909000 96 500 200 49000000 1 01 1 0 0 0 0 0196 119100 50 00000000000000000 Alkaline Phosphatase Labeling Kit NH 000 6 L3 L3 r3 ETE C3 C3 CDD L3 E3 Cd C3 C3 U ubt 300000000000000000000000000 888 Mw gt 50 000 000000000 0000006 MW 5 0000 000000
27. W Bernat M Johnson Anal Biochem 202 316 1992 F Schapper J T Goncalves M Oheim Eur Biophys J 32 635 2003 R R Duncan A Bergmann A M A Cousin D K Apps M y Shipston J Microsc 215 1 2004 Free radicals in biology and medicine 2nd ed Oxford Press 1988 C Schulze U Kleuker J Synchrotron Radiat 5 1085 1998 D J Brink N G van der Berg A J Botha Appl Opt 41 717 2002 Z Shi J J He S He J Opt Soc Am A Opt Image Sci Vis 21 1198 2004 J Swoger M Martinez Corral J Huisken E H Stelzer J Opt Soc Am A Opt Image Sci Vis 9 1910 2002 00000000000 010 http www nikonusa com poo http microscope olympus com 00000 http www zeiss com http www optronics co jp lex 0000000 http www joem or jp jis_maegaki htm 0000000000000 1 1 gt Home Page O 00100110 0105050 http Awww dojindo co jp msds index html DOJINDO Souci sm 3 2 PET 20274 FE EMM DIRA Material Satoty Data Sheet AN Xs RTE et SG mair ur ec Hz DE ces ENTRO E RETO AS 5 59 215 KUA YAMKUTA CCT T 774279 TEL 096 296 1515 FAX 0
28. 1 10 1 0000008000 color rendering 00000 40000 11 DOJIN News No 112 2004
29. 38 500 10 10 10 110000 10 Bu Cofee Vuen 31605 SADO 00000000000 8660 800 000 000 00 Cocoa units g 5 420 5 480 10 I 100 WST 18D00 3200 Table 3 SOSA of food samples extract with 80 ethanol produced in Kochi Prefecture Sample name SOSA units g dry Japanese name English name ESR WST 1 abaroncha Anaerobically treated tea 390 000 94 000 oishicha Post heating fermented tea 323 000 90 000 uwacha Mulberry tea leaf 86 400 21 000 hishitou Small sweet green pepper tr tr genmame Kidney bean tr tr hihoukuri Chestnut 352 32 900 Okura Okra 2 780 5 470 Derawea Delaware tr tr Kyuuri Cucumber tr tr Mlinitomato Cherry tomato tr tr ansouaroe Dehydrated aloe perrye tr tr Biwa kaniku Loquat pulp 297 tr Bwa kahi Loquat peel 1 470 1 460 syushi Loquat seed 2 580 3 860 umomo kaniku Plum pulp 811 1 610 umomo kahi Plum peel 3 110 13 500 Sumomo syushi Plum seed 36 000 22 100 tr ICso could not be determined under these conditions DOJ N News No 112 2004 OOOOO 00 Du DDD 1 0000000 0000000000000 37 411 1999 pogo0000000000000000000000pp 113 1279 0 1 1994 3 0000000 40800000037 328 1999 4 00000 Dojin News 96 1 2000 5 Okado Matsumoto and Fridovich Anal Bio
30. ADH alcohol dehydrogenase NOD NADH oxidase 51 00 p00000 5 1 0000 WST 10 000000000000 02000 11 1 0 0 0000 5 1 020000 1 1 10 000000000000 00 00 ep4o0g0000000000000 0000000000 11 HD Iu sobguu uguumuozagduuu u 5 0 0 00000000 111 1 1111 1 1 SOD 202 Y 2H gt H202 2 O 50 1 000000000000000 Guter O O O 1 2 H202 A O2N 1 2 O2 SOD H NA uric acid WST 1 Or OzN O2 HNS ON WA xanthine or N i hypoxanthine WST 1 formazan SOs Fig 8 Principle of the WST 1 method for estimation of SOD and superoxide anion scavenging activity SOSA WST 10000 SODOOOOOOOOOO Fig 8000 U 5091010000 02000 0 0 00 0 0 0 0 1 E 0 0
31. O 000000000 100 0000 020000 superoxide anion scavenging activity 90 5 5 0 00 80 F 00 0SOSAp0o00000000000000000000 e EI DO000000000000000000 00000000 2 601 zr D0000000000000000000000000000 ppgeopo 00000000 1 11 1111 111 481 1111 00 sosago oo 00000000000 09 cal sani aad al cuu 1111111010 11011111 0 001 0 01 0 1 1 10 100 1000 1 WsT100000000SOSAago 0000 Concentration of SOD U ml ugggugggaudggugggugdgaggagmgduuuusosA Fig 9 Inhibition curve of standard SOD preparation by the WST 1 method p0000000000000000000000000000 o000000000000000000000000 52 50000100 000020 00 000 0000000000 WST 1g 00090 1 SOD 0000XO000000000000000000000000 11 11111 1111 111 DO0O00000000000000000 WSt 1000000 O OSakudop 5 00000 000000000000 00020 0 000 000000000000 WST 100000 1111111 11111010 50 WST 1000000000 000 Hakkinen DO000000000000000 100000000 1 oo 00 0 0 00
32. Res Int 34 393 2001 21 0 48000000 pr 184 191 9908000000000 2004 22 A Sakudo D C Lee K Saeki Y Nakamura K Inoue Y Matsumoto S Itohara and T Onodera Biochem Biophys Res Commun 308 660 2003 23 J Hakkinen E Vehniainen and A Oikari Aquatic Toxicol 66 393 2004 24 T Ikegami Y Suzuki T Shimizu K Isono H Koseki and T Shirasawa Biochem Biophys Res Commun 296 729 2002 25 K H R Lin C C Weng H F Lo and J T Chen Plant Sci 167 355 2004 26 E J Woo J M Dunwell P W Goodenough A C Marvier and R W Pickersgill Nature Struc Biol 7 1036 2000 27 Y Shimasaki H Ohnishi S Matsuura and T Manaba Biochim Biophys Acta 15
33. gt 2 L3 L3 cJ r3 ES O e So ma E 3 3 oO 5 1 m E 055322 mg z Zo Ze 2 E Ed Gs c H L3 u Z E 2 op S s 5 a a a E 8 r3 cg 8 25 r3 9 E cec BE c m E A E r3 r3 o O gt C m O 25 I S 5 ELE S ki m O o E Fs r3 L3 SE 3 2 3 3 fa E SE a Eu r3 r3 EJ r3 EF ba ISO ES r3 0 gt ZOO m r3 r3 r3 r3 pud C3 cur Got 32 23 9 El A r3 5 3 5 a n c3 r3 L3 EX EC S pnm n OAT A Z 2222255555 E rm o SSES SBC ES E L3 3 3 60 090 601 r3 4 LO od 12 8 DOJAD News No 112 2004 0000 1 H Totsune M Nakano H Inaba Biochem Biophys Res Commun 194 1025 1993 R Creton L F Jaffe Biotechniques 31 1098 2001 M Masuko S Hosoi T Hayakawa FEMS Microbiol Lett 67 231 1991 Abramov J Gordon J Opt Soc Am 67 195 1977 P B Garland G H Moore Biochem J 183 561 1979 D Axelrod Methods Enzymol 361 1 2003 o0o0000 00000 54 245 2003 K Konig J Microsc 200 83 2000 J R Lakowicz H Szmacinski K Nowaczyk K
34. labelingg000000000000 000 00000000000 OD O O amber suppressor tRNA Mendel D Cornish V W and Schultz P G Annu 000000 Rev Biophys Biomol Struct 24 4350 1995m 000000000 Nemoto N Miyamoto Sato E and Yanagawa 00 00000000 H FEBS Lett 462 43 19990 0000 Griffin B A Adams S R and Tsien R Y Specific Covalent La beling of Recombinant Protein Molecules Inside Live Cells Science 281 269 1998 Guignet E G Hovius R and Vogel H Reversible Site selective Labeling of Membrane Proteins in Live Cells Nat Biotechol 22 440 2004 Keppler A Gendreizig S Gronemeyer T Pick H Vogel H and Johnsson K A General Method for the Covalent Labeling of Fusion Proteins with Small Molecules n Vivo Nat Biotechnol 21 86 2003 Keppler A Pick H Arrivoli C Vogel H and Johnsson K Labeling of Fusion Proteins with Synthetic Fluorophores in Live Cells Proc Natl Acad Sci 101 9955 2004 Giriat and Muir T W Protein Semi Synthesis in Living Cells J Am Chem Soc 125 7180 2003 Yeo D S Srinivasan R Uttamchandani M Chen G Y Zhu Q and Yao S Q Cell permeable Small Molecule Probes for Site spe cific Labeling of Proteins Chem Commun 2870 2003 1 DL Nucleostain DNA Damage Quantification K
35. 0 e NH Reactive ALPO 000000000000 2000000000 e Filtration Tube 0 00 0000 000000000000 00000 por 1 010 0000 0000000 30000000 NHz Reactive ALP 100 ug x 30 ALPO IgG Washing Buffer 4mlx 1 Reaction Buffer 200 ul x 11 Storage Buffer 4mlx 11 Fig 1 ALPD 67 Filtration Tube 30 Alkaline Phosphatase Labeling Kit NH J 0000000 pDooALPpo0000000000000NH gt Reactive ALP ooo 0000000ALPg00000000000 DODO 000000000000 0000000 19600 JUD po0p00000000000000 Filtration e 00 O Filtration Tube 9000000 CO CH 1 0 ig 1 PO UU UU un DODODOO Alkaline Phosphatase Labeling Kit NH2 3 samples 21 000 LK12 OO O O O Filtration Tube 800000 OOOOOOOOOOOO C L3 E C3 C3 lt 0 LI ELE E s E3 EJ E rrr 0 go 1 D pgg0080 00 0000000000000 1 go 8 DOJAD News No 112 2004 Alkaline Phosphatase Labeling Kit SH K 0000 sa 0000 Mw gt 50 000 000000000 0000 IgG lt T Mw lt 5 0000 0000000 e SH Reactive ALPO 0 0 0 000000000000000
36. 00 0000000000000 ssgogog0 000000000000 30000000 SH Reactive Biotin 30 87 300 00 Reducing Agent 30 I 100 0 Reaction Buffer 1 10 WS Buffer 4mlx 10 0000000000 Filtration Tube 3n Ti Reactive Biotin Biotin Labeling kt GH SH 0000000000000 AC 1 11101100000000000 SH Re active Biotin 0000000 6 S SO 1 1 COOC 37 300 0 0 5 ai 2 5 3 c 0 0 6 0 1 D U 1 1 uu ng 0000 Filtration 111111 1 1 5 1 ot CO 5 lt Y o 0 0000 500 200 490 0 D 0000 0000 00000000 896 UU UU a Z H oo000000 l60000 8 DOJI News No 112 2004 L 1 1 Oyster 5
37. 00000 Dado gg gg O DO Doo 0000000000000 L DO UUUIUUD 0000000000000 ALPD Alkaline Phosphatase Labeling Kit SH DODoO go ag 0 o 0000000000 3 samples 21 000 LK13 News No 112 2004 96 111 Q amp A TR o a 111 ana Alkaline Phosphatase Labeling Kit NH Q7 80000000085500000000000000 Alkaline Phosphatase Labeling Kit SH 00000000 A7 popop00 00000000000 0S SoO SHO 0 0000000000 Peroxidase Labeling Kit O O O 000000 Reducing Agent 00000000000 11 1 1 1111111 111 11111 111 Peroxidase Alkaline Phosphatase 000000000 08 pogo aca 00000000 0000000000 Du Q1 01 1 20 1 Ae 00000 1 Ai 0000000000000000 Alkaline Phosphatase Dg 00000 000000000 0000000000 Labeling Kit NH j SHO 0000 S S000000 o gogo ogg 000000 Alkaline Phosphatase Labeling Kit SH 1 O O U Du Q9 Reactive ALP NH2 SH O 0 0 buffer 0000000 0000 Q2 igc00000000000000000000000 A9 DQDO00000000000000000000 A2 0000 50 0000000005 0000000000000 Reactive ALPO 0 00 0 000 000000000000 111111 1 000 5 00011100 00 00 000000 000 11 1111
38. 000000 10 1 00gwsT 19 O 01 00100 00 0010017 0000 un O DD r 0 9994 16 n 0 O 0 Fig 5 00 O 000 1 1 5009 0 0000 0000 SOO 3000 0000 0000 1 O 00000 XON 11 1000000000 Hypoxanthine 0 9 Y rormazan Y 1 038X 0 00016 Cie HO 4 Na 2oz WST E Al gt 049994 i n 16 Xanthine o Oz 202 Formazan 5 Pe X E 1 02 H20 Catalase H202 l 207 WST 1 5 0 3 Uric acid I 0 1 1 IV XO Catalase 0 0 3 0 6 0 9 i 2 Carrier P 1 D gt Waste FIA method mM Fig 5 Correlation between the conventional HPLC method and the FIA method for the determination of hypoxanthine in fish samples X and Y indicate the concentration mM obtained by the present FIA method and the HPLC method respectively WST 1 Hypoxanthine or Water Fig 4 Principle and FIA manifold of hypoxanthine assay P pump IV injection valve D detector DOJ N News No 112 2004 Absorbance at 438 nm 0 0 0 01 0 02 0 03 0 04 0 05 NADH concentration mM Fig 6 Calibration curve of NADH based on the detection of O2 formed in the oxidation of NADH
39. 000000 phosphorescencegg O O O DD DODO 0000 00000000000 UU 1 0 000000000000000 0 1 LU retina 0000000000 LU rod 681 gg dBg8 000 000000000000 000000000 00000000 0 00 1 Ix 000000100000 1 680 Wm99 0000000 090800000000 0 00000000 0 1 0 0 D LED light emitting diode 50 0 DD HIDO High intensity dis 1 charge lampp 1000 0000000 100000000000 pogo0doed000000000 000000000000 000 300000 ag a ooo 000000000000 D reflection 0 0 absorption T O O transmission OOO 000 31 0000 11 0 0 0 09006 68 1 000000000
40. 00000000 5580 0 0 0 0000000000 00 0000000000000000 0 0 00 0 00 0000 00 00 0 00005 1 1000000000 DO D 0000000000000 10 0 0 0 0 0 60 Oe DD D NADPH 080 0000000000000 00 0 00 0 0 00000000000 0000000000000 0 1 0000000000 0000000 00000000000 0 00 0 000000 0000 60000000 0 0000 000000000000000000 0 0 0 8 00 0 8 70 00000000000 1 000000000000 00000 00000000000 111101111111111 0000000000 DOJM News No 112 2004
41. 000000000000000 NHeReac tive Bioting 0 0 Filtration Tube 0000000000 0000001600004 0D0000000000000000000000000000 ia 00000000000000000000000 10000000000000000 son 20009008 Biotinylation Kit Sulfo OSu 0000000000000000 Biotinylation Kit Sulfo 000000000000000000000 Biotin ACs 2 ulto 0s up poo 00000000000000000000 000000000000000000000 100000 00 0 0 0 NaHCO 0000000 0000000000000000 8500000000 000 Biotinylation Kit Sulfo OSu 0 10000000 10 5889800000 COOCO0OOCOOCOOCOOCOO0OCOOCO1400000000 10 mg x400000000000000 4000000000000 D D DD D D Biotin ACs zSulfo OSu D 000 000000 000000000000000000 gt gt 0 Biotin ACs 2 Sulfo OSu 10 mg x 40 D NaHCOsQ 00000 40 pj PBS I O 40 000000000 40 000000000 80 0 0 ooomoo 0000000 10 0 ooomoo 0000000 Biotin Labeling Kit NH2 Biotinylation Kit Sulfo OSu 3 samples 12 000 LK03 set 35 800 BK01 News No 112 2004 96 111 1 Biotin Labeling Kit SH Ar 00 6 q E 0000 IgG e3s0000000000000000000 e SH Reactive Biotind 0 0000000000000000 UU 1100000000 00 00 000000000 0000000000 0 e Filtration Tube 00000000000000 00000 Reducing Agent 00000 4 1 80000000
42. 1 Q10 90000 butter 0 0 0 450 n 0000 OOOOOOOOOOOOOOO Q3 00000 8960 0 0 0000000000000 0 A10 D00000000000000000 A3 00000 9G000 500 2009000000000 10 u90 9 8 Q11 096 1 O U Alkaline Phosphataser 11 11111 1111111 111111 1 A11 9011010000 5800 IgG 1000 Q4 DO00000000000000000000000 000000 10 2000 Alkaline Phosphatase O U A4 Dual OO 00 0 Filtration Tube 4000000000000 O O Alkaline Phosphatase 00000000000 1111 1 0000000 0 5 n DO O O Filtration 1 UN 500 1 1 11 0 0 0000 9 D 200 90000000000 0 0 O Filtration 0 0 0 0 0 00000 0000 111111 1111 11 11 WU Q12 200 u90 000000mg00000000000000 500 200 g0 111111 1 111 1 1 11 111111 111
43. 1 2 Ou 252525235550 29205725085 d CH cO 9 5 8 o Cm CDI Bu a a G E BABE EE ES D e N I O Rei ro 5 S OER m 6 G OO gt 8 9 3 amoaoogoruugOooudo z ox t ouumnuguogguouuurn dez d 5 a 025005070500 5 5 e 222525322025 a 2 C2 C3 Ss SES 5 T 10505567 ERE 5 DODO 5 0 DCD DD 4 ir EE CO COse EI 0 CC lt CC COOC 2 E 7 H ra EL 0008 EE H E 3 E x gt 0 El J J oooh 0 CDD oan Pr rns COOC Soot GOGO EJ C3 DE La 63 3 2 2 35 Ei E E EX EX Ed ET EB r Lir 5353 53 EX py 5 Eu ES CDD a200 o 9 2985525550 3 3 3 53 53 53 2225532527 2 0 SOs 2 3 ELE EE EE E en 5
44. 2 2004 OE L3 50000 nano o 4 a g 4 4 3 53 UU 10 50 60 70 I 2 O 17 10 umol l 11111 1 1 1 z o EA CH C3 rr r3 3 E su E cd 2 gt Z gt a i oL 51 4 c 2 n3 gt m lt 2 N a D o o 2 5 E L3 r3 EJ C3 Hi pc rm ZE C3 r3 p nr d pn 0 EJ p rp EJ ELE Eng E3 EE Lr COD 20000 g IJ 1 L2 L3 p LJ LJ LC CJ LIII C LI a a E 50003 0000 0000 c3 p c rne mn DUG luu a r3 a E BBS x EE CC I 000000000000000inteing 0000 in vivo
45. 2000 D 1 1 WST 10 00 10 0 0 gt ga El 2 2 2 070 80 00 0 SOD jo 0 0 0 0 0 0 0 L3 p4 CJ x LJ CJ p4 0 0 0 0 1 o C p a 0 DL U D pH 10 20 DOJM News No 112 2004
46. 56 Labeling Kit NH2 Oyster 656 Labeling Kit NH noo 50 200 90 210000 30000000000000 Oyster dye Denovo biolabelst 0000000000 000000000200 000000000000000 0000000008 dayepogo 000000000000 000000000000 00000 0000000000 DO 0000 Fluorescein Labeling Kit NH2 gt 00 lt gt 00 lt 00000 0200000000 020000000000000 e Filtration Tueeggp000000000000000000 e Filtration 00000 00000 lt gt 30 Reactive Fluorescein WS Buffer 4mlx 11 Reaction Buffer 500 ul x 10 lt 00000 gt Filtration Tube 30 NH2 Reactive Oyster dye 30 WS Buffer 4 ml x 1 Reaction Buffer 500 ul x 10 Filtration Tube 30 Oyster 556dye 0 D O Oyster 656dye 0000000 R f 1 E 035 R N Ra 0S R N Ra L ap R N Kt 1 RR Ri RA sos K CHa s Q 2 0 0 Amine Reacti
47. 71 245 2002 28 T Finkel and N J Holbrook Nature 408 239 2000 29 K Suetsuna and H Ukeda J Nutr Biochem 11 128 2000 30 000000000000 O U 65 1096 1999 3 49 25 2002 32 0 0 49 679 2002 a3 O A O 80 392 2003 34 09 2003 35 N Tsuji N Hirayanagi M Okada H Miyasaka K Hirata M H Zenk and K Miyamoto Biochem Biophys Res Commun 293 653 2002 poo 11 Hiroyuki Ukeda o 000 783 8502 000000 200 Tell Fax 088 864 5189 E mail hukeda cc kochi u ac jp 1 D 111 1 1 1111 111111 11 1 1 1 1B ED EL UL 111 H 1 E DI SOD Assay Kit WST lt 00 gt e100 Ss0Dpp a 000000000000 eut uuu uuu 111111 SOD Assay Kit WSTO 00 0000000000000090 00000 WST 10 O U LI 180000000WST 1 xanthine oxidase XO 000 0 100 0 0 000 000000000000 960 1111 1 111 U
48. 96 206 1525 0000000 770 100 1400 02700000 120 800 D 000110000 1 1111110110 UU 160000 000000 0000000 0000000 DOJM News No 112 2004 Topics on Chemistry 000000 In viven 10 00000 OOO ol SN R H NTA I R 503 NTA II R Fig 1NTADODO DODO 0000000000000 00000000000 0000000 Ka lt 10 prof gg 000000000000 0 0 00 0 00 11 0 0 0 000000 1 DU 46 1890000 0 0 0 O Guignetg 0 0000NTAD 00 Fig 100 O 88 89 NTAD 1110100000000 3000000000 Hiss 400000000000 0 011 0 His Tag O 0 O U UD 0000000000000 DODO 0000000000000 000000000000
49. I 0000000 0000000 SOD Assay Kit WST 500 tests 19 000 S311 DOJI News No 112 2004 2 2 Dd 000000000000 00000002100 0 DD S Z X 100000 0080000000000 1111111011 0000 0000 unu 11 1 0000 0000 111 1 L3 a p gm rm a 0 L3 C3 C3 L3 rJ C3 C3 2 r CJ C3 C3 C3 O C3 C3 EJ E EJ EJ E EJ EJ LE EI 1 go 1 D D 1 1 D EI UU 1 go D 1 1 UUUU 000 E 000 0000 electromagnetic wave 00000 holz 18520 Mog00000000000000 OOOO 111 2800000000000 O wavelength 0000000000000 0000000Fi9 100000 2 2 o E I 3 DO 0000000000 0000000000 00000000000 ultraviolet UVIT OOOO aw
50. N Ac Lys Aldehyde alone Glyceraldehyde 19 1 8 51 10 7 5 36 Glycolaldehyde 8 70 1 43 7 12 4 28 Threose 39 1 4 46 13 4 0 713 Erythrose 48 0 16 2 13 8 4 84 Xylose 1 61 0 203 0 356 nd nd means fet detected
51. chem 298 337 2001 6 H Ukeda S Maeda T Ishii and M Sawamura Anal Biochem 251 206 1997 7 H Zhao S Kalivendi H Zhang J Joseph K Nithipatikom J Vasquez Vivar and B Kalyanaraman Free Rad Biol Med 34 1359 2003 8 M A Barbacanne J P Souchard B Darblade J P Iliou F Nepveu B Pipy F Bayard and J F Arnal Free Rad Biol Med 29 388 2000 9 00000 Dojin News 82 10 1996 10 H Ukeda D Kawana S Maeda and M Sawamura Biosci Biotech Biochem 63 485 1999 11 T Oritani N Fukuhara T Okajima F Kitamura and T Ohsaka Inorg Chim Acta 357 436 2004 12 A S Tan and M V Berridge J Immunol Methods 238 59 2000 1 14 T Tsukatani T Oba H Ukeda and K Matsumoto Anal Sci 19 659 2003 15 H Ukeda Y Hasegawa Y Harada and M Sawamura Biosci Biotech Biochem 66 36 2002 16 H Sekido T Suzuki T Jomori M Takeuchi C Yabe Nishimura and S Yagihashi Biochem Biophys Res Commun 320 241 2004 17 H Ukeda T Shimamura M Tsubouchi Y Harada Y Nakai and M Sawamura Anal Sci 18 1151 2002 18 H Iwata H Ukeda T Maruyama T Fujino and M Sawamura Biochem Biophys Res Commun 321 700 2004 19 A K Sarker H Ukeda D Kawana and M Sawamura Anal Sci 15 1141 1999 20 A K Sarker H Ukeda D Kawana and M Sawamura Food
52. it AP Site Counting Nucleostain DNA Damage Quantification Kit AP Site Count ing 0 0 0 0 000 000000000000 DNADDDD AP site LO 0 00 00 00000 00 0000 000 0000 HRP streptavidin p O 0 0 0 00 0 00 00 0 0 0 0 00 0 0 pa40 00000 000000000 00000000000 1 01 1 http www dojindo co jp backnumber news ddak html DO HRP streptavidin j500ux 1 000 0025ux1 DODO 1 1 1 1 1 1 00 DD 2 AP site 0000 HRP streptavidin 400 ul O PBST 19 6 mig 5008000000 HRP streptaviding 0000000 DUU O 0 HRP streptavidin PBST 4 0000 1 0 00 HRP streptavidin 0 O 0 HRP streptavidin 10 ul PBST 40 ml Va ETE ET seus 7 00000 HRP streptavidind 40 000000000000 1111111 11111111 1101 101 1 1 11 111 11 1 1 1 001 500 0 0 0 0 HRP streptaviding 00 1 4 0000 0 0 0 HRP streptavidin 00000000
53. ming MnO 1 000 0 8 0 17 0000 Geming 0 0 000000 0 0 0000000090 000000000000 Shimasakig WST 10 000 11111111111 1 og ag 0000000 DOJ N News No 112 2004
54. ulated emission of radiation laser 1 0000000 1101 00 000000000000000 Du 0 00 360 nm 400 0 0 O 760 O 00 bright O a 0 nm 830 nm 000000000 O 00 prism O D white light O O U O 100090 D Fig LL UO B ELO 1 U 1111 1 1 111 2 17020 Newton 7 00 spectrum D coherent OOO DOD Uu 0000 0000000090 1 lens O O 11111 1 1 23 000000 Fig 30 234 0000000090 H OUUU 1 0000000000000 0 0 bioluminescence OOO 111111 O luciferin O O 0 chemiluminescence D Fig 4 ug DU 00000 E 1 DU 100000000 photon counting 0000000 ATP 0000000 1 000 2 3 2 00090 Leeuwenhoek 170 Fig 4 Chemiluminescence from bamboo shoot cut Each spot reflects a 0000000090 001900 Edison single photon derived from excited tyrosine 10 DOJ N News No 112 2004 fluorescence 0000000 33 0000000090 2 3 3 000
55. ve Oyster 6 Amine Reactive Oyster 6 mE 00 00000 Amine Reactive Fluorescein 0 0 00000 0 00000 0 0 00000 0 00000 Fluorescence Intensity arbitrary Fluorescence Intensity arbitrary Fluorescence Intensity arbitrary 400 450 500 550 600 650 450 500 550 600 650 700 550 600 650 700 750 800 Wavelength nm Wavelength nm Wavelength nm Fig 1 Fluorescein Labeling Kit O Fig 2 Oyster 556 Labeling Kit NHa O O O Fig 3 Oyster 656 Labeling Kit NH20 0 O O 21 DOM News No 112 2004 1508100000 0 0 00 2004 110 26 9 30 17 300 0 O 9 000 0 17 6 1 300 Li LI Ll B 1
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