Lڠاอ΅ΛΠ 87 ήυΛ΅ϋΈဣס Chemi-Lumi OneBlocking One Chemi-Lumi One 䊒䊨䊁䉥䊷䊛⎇ⓥ䈏⋓䉖䈮ዷ㐿䈘䉏䈩䈇䉎ᣣ䇮᛫ේ᛫ᔕ䉕↪䈚䈢∉ቇ⊛ᚻᴺ䈮䉋䉎䉺䊮䊌䉪⾰䈱․⇣⊛ᬌ䊶หቯ䈲䇮䊋䉟䉥䊁 䉪䊉䊨䉳䊷ಽ㊁䈮䈍䈇䈩ၮ⋚䈎䈧ᔅⷐਇนᰳ䈭ᛛⴚ䈫䈭䈦䈩䈇䉁䈜䇯ਛ䈪䉅䇮㔚᳇ᵒേᓟ䈱䉺䊮䊌䉪⾰䉕䊜䊮䊑䊧䊮䈮ォ౮䈚䈢ᓟ䇮X✢䊐䉞 䊦䊛䈮ኒ⌕䈘䈞䇮ᓸ㊂䉺䊮䊌䉪⾰䉕․⇣⊛䈎䈧㜞ᗵᐲ䈮ᬌ䈜䉎ൻቇ⊒శᬌᛛⴚ䈲䇮ᦨ㜞ፄ䈱ᬌᚻᲑ䈪䈅䉍䇮ᦨ䉅᳢↪ᕈ䈱㜞䈇ᬌ ᛛⴚ䈪䈜䇯ㄭᐕ䇮⍴ኼ䈪䈅䈦䈢ൻቇ⊒శ䈏ᛛⴚᡷ⦟䈘䉏㐳ኼൻ䈚䇮䊑䊨䉾䊁䉞䊮䉫ᬌ䈭䈬䈮ᔕ↪䊶᳢↪䈘䉏䈩䈇䉁䈜䇯䈖䈖䈪䈲ᒰ␠䈪 ⽼ᄁਛ䈱ൻቇ⊒శᬌ䉨䉾䊃䉕⚫䈚䉁䈜䇯 ․˴㐳 ٨ჺ᧓Ὁ᭗ज़ࡇ౨Јỉ҄ܖႆήỿἕἚ ٨˯ẟἢἕἁἂἻỸὅἛỂᅵễἙὊἑ౨Ј ຠ⺑ ᬌේℂ㩿ㅢᏱᬌ㪀 ᬌේℂ㩿㜞ᗵᐲᬌ㪀 䋨䉰䊮䊄䉟䉾䉼ᴺ䋩 䋨䉝䊎䉳䊮㪄䊎䉥䉼䊮♽↪䋩 ㆊ㉄ൻ‛ Luminol ⊒శ ㆊ㉄ൻ‛ Luminol ൻቇ⊒శ䈱ᔕ䊜䉦䊆䉵䊛 ⊒శ ࠛࡦࡂࡦࠨ ࠛࡦࡂࡦࠨ 21& ࡆࠝ࠴ࡦ21& ᰴ᛫ ᰴ᛫ 㧔21&ᮡ⼂᛫㧕 ᰴ᛫ 㧔ࡆࠝ࠴ࡦ᛫㧕 ࠕࡆࠫࡦ ᰴ᛫ ࠲ࡦࡄࠢ⾰ ࠲ࡦࡄࠢ⾰ ⊒శ 䊦䊚䊉䊷䊦䋨Luminol䋩ᔕ䈫䈲䇮䊦䊚䊉䊷䊦䈫䈇䈉ൻว‛䈏㉄ൻ䈘䉏䈩䇮3-䉝䊚䊉䊐䉺䊦㉄䈮䈭䉎ᔕ䈪䈜䇯 䊕䊦䉥䉨䉲䉻䊷䉷䈲䇮ㆊ㉄ൻ᳓⚛䈫䊦䊚䊉䊷䊦䈱ᔕ䉕⸅ᇦ䈚䇮㕍⦡䈱Ⱟశ‛⾰3-䉝䊚䊉䊐䉺䊦㉄䉕↢ᚑ䈚䉁䈜䇯 ↪ᣇᴺ ㅢᏱᬌᴺ䊶㜞ᗵᐲᬌᴺ⇛࿑ 㔚᳇ᵒേ䊶䊑䊨䉾䊁䉞䊮䉫 䊑䊨䉾䉨䊮䉫 (BlockingOne↪) 1ᰴ᛫ᔕ ㅢᏱᬌᴺ 㜞ᗵᐲᬌᴺ PODᮡ⼂᛫ᔕ 䊎䉥䉼䊮ᮡ⼂᛫ᔕ 䉴䊃䊧䊒䊃ABC 䊕䊦䉥䉨䉲䉻䊷䉷䉨䉾䊃ᔕ Chemi-LumiOne ⊒శ↪ᷙวᶧᔕ ᗵశ䊶䊶ቯ⌕ ↪ᣇᴺ 1 㽲 䉰䊮䊒䊦䇮䊎䉥䉼䊮ᮡ⼂䉺䊮䊌䉪⾰䊙䊷䉦䊷* (ຠ⇟ภ䋺02886-44)䋨䉅䈚䈒䈲㕖䊎䉥䉼䊮ᮡ⼂ 䉺䊮䊌䉪⾰䊙䊷䉦䊷䋩䉕㔚᳇ᵒേᓟ䇮䉭䊦䉕⤑䈮ォ౮䈚䇮0.05䋦 Tween 20TBS(t-TBS)䉕↪䈇 䈩⤑䉕ᵞᵺ䈚䉁䈜䇯(5ಽ㑆㬍2࿁) 㽳 䊑䊨䉾䉨䊮䉫䊪䊮*3 (ຠ⇟ภ䋺03953-95)䉕↪䈇䈩45ಽ㑆 ቶ᷷䈪䊑䊨䉾䉨䊮䉫ಣℂ䉕ⴕ䈇䉁䈜䇯 㽴 t-TBS䉕↪䈇䈩⤑䉕ᵞᵺ䈚䉁䈜䇯(5ಽ㑆㬍2࿁) 㽵 1ᰴ᛫䉕ᔕ䈘䈞䉁䈜䇯(ቶ᷷䇮1ᤨ㑆) 㽶 t-TBS䉕↪䈇䈩⤑䉕ᵞᵺ䈚䉁䈜䇯(5ಽ㑆㬍2࿁) 2 㽷 䊎䉥䉼䊮ᮡ⼂᛫ (2ᰴ᛫)* (䉅䈚䈒䈲PODᮡ⼂᛫)䉕ᔕ䈘䈞䉁䈜䇯(ቶ᷷1ᤨ㑆) 2 㽸 t-TBS䉕↪䈇䈩⤑䉕ᵞᵺ䈚䉁䈜䇯(5ಽ㑆㬍2࿁)* 㽹 䉴䊃䊧䊒䊃ABC䊕䊦䉥䉨䉲䉻䊷䉷䉨䉾䊃 (ຠ⇟ภ䋺30462-30)䈱Working Solution䉕⺞䈚䇮 2 t-TBS䈪10Ꮧ㉼䈚䈢ṁᶧ䉕ᔕ䈘䈞䉁䈜䇯* 㽺 t-TBS䉕↪䈇䈩⤑䉕ᵞᵺ䈚䉁䈜䇯(5ಽ㑆㬍2࿁) 㽻 Solution A䈫Solution B䉕1䋺1䈱ഀว䈪ᷙว䈚䉁䈜䇯 (ᷙวṁᶧ䈲䊑䊨䉾䊁䉞䊮䉫䈚䈢⤑䈮ኻ䈚 䈩⚂ 0.125ml/cm2䈮䈭䉎㊂䉕↪䈚䉁䈜䇯) 㽼 䊒䊤䉴䉼䉾䉪䊤䉾䊒䊐䉞䊦䊛䉕䈮ᢝ䈐䇮䈠䈱䈮䊑䊨䉾䊁䉞䊮䉫⤑䈱䉺䊮䊌䉪⾰ๆ⌕ 䈏䈮䈭䉎䉋䈉䈮⟎䈐䉁䈜䇯 㽽 0.125ml/cm2䈱⊒శ↪ᷙวᶧ䉕⤑䈮タ䈞䇮1ಽ㑆ᔕ䈘䈞䉁䈜䇯 㽾 ⤑䈱┵䉕䉨䊛䊪䉟䊒䈮⌕䈔䉎䈭䈬䉕䈚䈩䇮ಽ䈭ᷙวᶧ䉕ข䉍㒰䈐䇮⤑䉕චಽ䈮൮䉄䉎⒟ᐲ䈱 ᄢ䈐䈘䈱ᣂ䈚䈇䊒䊤䉴䉼䉾䉪䊤䉾䊒䊐䉞䊦䊛䈮䉺䊮䊌䉪⾰ๆ⌕䈱㕙䈏ਅ䈮䈭䉎䉋䈉䈮 䊑䊨䉾䊁䉞䊮䉫⤑䉕⟎䈐䉁䈜䇯 (䈖䈱ᤨ䇮⤑䈫䊤䉾䊒䊐䉞䊦䊛䈱㑆䈮᳇ᵃ䈏䉌䈭䈇䉋䈉䇮 䉁䈢䇮䊤䉾䊒䊐䉞䊦䊛䈱᛬䉏䉇䉲䊪䈏㕙䈮᧪䈭䈇᭽䈮ᵈᗧ䈚䈩䈒䈣䈘䈇䇯) 㽿 䊤䉾䊒䊐䉞䊦䊛䈪䊑䊨䉾䊁䉞䊮䉫⤑䉕൮䉂䇮䉺䊮䊌䉪⾰ๆ⌕䈱㕙䉕䈮䈚䈩䊐䉞䊦䊛䉦䉶䉾 䊃䈮⟎䈐䇮䊐䉞䊦䊛䉦䉶䉾䊃䉕ᥧቶ䈮⟎䈐䉁䈜䇯 㾀 ᥧቶౝ䈪䇮䊤䉾䊒䊐䉞䊦䊛䈪൮䉖䈣䊑䊨䉾䊁䉞䊮䉫⤑䈱䈮X✢䊐䉞䊦䊛䉕⟎䈐䇮䊐䉞䊦䊛䉦 䉶䉾䊃䉕㐽䉄䇮3ಽ㑆ᗵశ䈘䈞䉁䈜䇯 㾁 X✢䊐䉞䊦䊛䉕ᶧ䈮ẃ䈔䇮䈏⏕䈪䈐䉎䉁䈪䈚䉁䈜䇯 㾂 X✢䊐䉞䊦䊛䉕ቯ⌕ᶧ䈮ẃ䈔䇮䉕ቯ⌕䈘䈞䉁䈜䇯 㾃 X✢䊐䉞䊦䊛䉕ᥧቶ䈎䉌䈚䇮⚿ᨐ䉕⏕䈚䉁䈜䇯 *1) 䊎䉥䉼䊮ᮡ⼂䊙䊷䉦䊷䉕↪䈜䉎႐ว䇮䉴䊃䊧䊒䊃ABC♽⹜⮎䈱↪䈲ᔅ㗇䈪䈜䇯 *2) ㍈ᢅ䈭ᬌᴺ䈫䈚䈩䈖䈱ᠲ䉕䈗ផᅑ⥌䈚䉁䈜䇯 㵘㵘PODᮡ⼂2ᰴ᛫䋨ㅢᏱ䈱䉰䊮䊄䉟䉾䉼ᴺ䋩䉕↪䈜䉎႐ว䈲䇮䈖䈱ᠲ䈱ᔅⷐ䈲䈅䉍䉁䈞䉖䇯 *3) 䊑䊨䉾䉨䊮䉫䊪䊮䈲䇮䉝䊎䉳䊮䊶䊎䉥䉼䊮ᔕ䉕↪䈚䈢ᬌ♽䈪䈲䇮ᧄᶧ䈱䉡䉲↱᧪䉺䊮䊌䉪⾰䈫 Ꮕᔕ䈜䉎น⢻ᕈ䈏䈅䉍䉁䈜䇯䉁䈢ᧄᶧ䈮䈘䉏䈩䈇䉎䉺䊮䊌䉪⾰䈲䇮♖䉺䊮䊌䉪⾰䉕↪䈚䈩 䈇䉁䈜䈏䇮᷹ቯ♽䈱᛫䉇㉂⚛ᮡ⼂᛫䈭䈬䈫Ꮕᔕ䈜䉎น⢻ᕈ䈏䈅䉍䉁䈜䇯↪䈮వ┙䈦䈩䇮 Ꮕᔕ䈱ή䉕䈗⏕䈒䈣䈘䈇䇯 ↪ Chemi-Lumi One䉕↪䈇䈢ᬌ⚿ᨐ Ԙ˴ԙԚԛ %Ԝԝ 䊧䊷䊮㽲䌾㽷㩷䋺㵘2㱘g / 䊧䊷䊮 C 䋺㵘䉮䊮䊃䊨䊷䊦 : 10 㱘g or 20 㱘g of Drosophila Schneider-2 cell lysate ⤑ : PVDF⤑ ࡉࡠ࠶ࠠࡦࠣ : 5% dry milk / TBS, 30 min, RT ৻ᰴ᛫ : anti-Akt H136 (1/1000, Santa Cruz) / 0.25% BSA / TBS, 10 h, 4㷄 ᵞᵺ : 0.5% Tween-20 / TBS, 10 min x 3, RT ੑᰴ᛫ : anti-rabbit IgG, HRP-conjugated (1/2000) / 0.5% Tween-20 / TBS, 30 min, RT ᵞᵺ : 0.5% Tween-20 / TBS, 10 min x 5, RT 㔺శ : 1 min ࠨࡦࡊ࡞ ࠺࠲ߏឭଏ㧦᧲੩ㇺ⥃ᐥකቇ✚ว⎇ⓥᚲ⮎ℂ⎇ⓥㇱ㐷 ˴˴˴˴˴˴˴೨ựᦺᒾవ↢ ࠨࡦࡊ࡞ ⤑ ࡉࡠ࠶ࠠࡦࠣ ৻ᰴ᛫ ੑᰴ᛫ 㔺శ : ࡑ࠙ࠬ⣖ᶧ : ࠾࠻ࡠ࡞ࡠࠬ : 5% ࠬࠠࡓࡒ࡞ࠢ/ 0.05% PBST :anti-APP,Ctm (1/3,000, overnight, 4͠) :HRP-conjugated Anti Rabbit (1/5,000, 1hr, RT) :3 min ࠺࠲ߏឭଏ㧦ℂൻቇ⎇ⓥᚲ˴ ⣖⑼ቇ✚ว⎇ⓥࡦ࠲ ˴˴ ↪ 䉝䊎䉳䊮㪄䊎䉥䉼䊮䉕↪䈇䈢㜞ᗵᐲᬌ ԘԙԚ ԛ 㽲䊎䉥䉼䊮ᮡ⼂䊙䊷䉦䊷 㽳䊙䉡䉴ⴊᷡ1000Ꮧ㉼:0.18㱘g/䊧䊷䊮 㽴䊙䉡䉴ⴊᷡ1000Ꮧ㉼:0.36㱘g/䊧䊷䊮 㽵䊙䉡䉴ⴊᷡ1000Ꮧ㉼:0.72㱘g/䊧䊷䊮 䊎䉥䉼䊮ᮡ⼂᛫䊙䉡䉴IgG᛫䈫ᔕᓟ䇮䉴䊃䊧䊒䊃ABC䊕䊦䉥䉨䉲䉻䊷䉷䉨䉾䊃䈶 Chemi-Lumi One䋨㔺శᤨ㑆䋺3 min䋩䉕↪䈇ᬌ䈚䉁䈚䈢䇯Ꮐ࿑䈱䊂䊷䉺䈎䉌䇮䉝䊎䉳䊮䊎䉥䉼䊮䉕↪䈇䈢ᬌ♽䈮䉋䉍䇮ᓸ㊂䈱䉺䊮䊌䉪⾰䉕㍈ᢅ䈮ᬌ䈜䉎䈏น⢻䈪䈜䇯 䊥䊒䊨䊷䊎䊮䉫䊂䊷䉺 ઁ␠ຠ B Chemi-Lumi One 㽲 䊍䊃ⴊᷡ (1000Ꮧ㉼), 䊙䉡䉴ⴊᷡ(1000Ꮧ㉼)䈱ᷙว‛ 2.5, 5.0, 10.0 㱘l 䉕SDS䋭PAGE 㽳 䉶䊚䊄䊤䉟䊑䊨䉾䊁䉞䊮䉫 (PVDF⤑) 㽴 䊑䊨䉾䉨䊮䉫 (Blocking One) 㽵 ᛫䊙䉡䉴IgG᛫ (PODᮡ⼂)䈪ᔕ 㽶 ൻቇ⊒శᬌ 㽷 䊥䊒䊨䊷䊎䊮䉫 [Tris✭ⴣᶧ(67.5 mM, pH 6.8)(2䋦 SDS, 100 mM 2-䊜䊦䉦䊒䊃 㵘䉣䉺䊉䊷䊦) ]70㷄, 45min 㽸 䊑䊨䉾䉨䊮䉫 (Blocking One) 㽹 ᛫䊍䊃䉝䊦䊑䊚䊮᛫ (PODᮡ⼂)䈪ᔕ 㽺 ൻቇ⊒శᬌ ࿑䈱䊂䊷䉺䈎䉌䇮Chemi-Lumi One䈲䊥䊒䊨䊷䊎䊮䉫䉕ⴕ䈦䈩䉅↪น⢻䈪䈅䉍, 䉁䈢ઁ␠ຠ䈫㆖⦡䈱䈭䈇ᕈ⢻䉕␜䈜䈏䇮ታ⸽䈘䉏䈩䈇䉁䈜䇯 ຠౝኈ Solution A : 䊦䊚䊉䊷䊦ṁᶧ 250 ml Solution B : ㆊ㉄ൻ‛ṁᶧ 250 ml 䊶ᧄ䉨䉾䊃䈮䈲⣣㘩ᕈ䈱‛⾰䈏䉁䉏䈩䈇䉁䈜䇯䈗↪䈱㓙䈮䈲ᚻⴼ䇮㒐⼔⌒㏜䉕⌕↪䈚䈩䈒䈣䈘䈇䇯 䊶⹜⮎䉕⺞䈜䉎㓙䈮䈲䇮ᔅ䈝ቶ᷷䈮ᚯ䈚䈩䈎䉌↉䉕㐿ኽ䈚䈩䈒䈣䈘䈇䇯 䊶䊑䊨䉾䊁䉞䊮䉫⤑䈲ᚻⴼ⌕↪䉁䈢䈲䊏䊮䉶䉾䊃䉕↪䈇䈩ข䉍ᛒ䈇䈒䈣䈘䈇䇯 䊶⤑䉕䊤䉾䊒䊐䉞䊦䊛䈪൮䉃㓙䇮⤑䈫䊤䉾䊒䊐䉞䊦䊛䈱㑆䈮᳇ᵃ䈏䉌䈭䈇䉋䈉䇮䉁䈢䊤䉾䊒䊐䉞䊦䊛䈱᛬䉏䉇 㩷㩷䉲䊪䈏㕙䈮᧪䈭䈇᭽䈮ᵈᗧ䈚䈩䈒䈣䈘䈇䇯 ଔᩰ ຠฬ Chemi-Lumi One ⷙᩰ ⾂ᴺ ຠ⇟ภ ኈ㊂ ଔᩰ SP ಄⬿ 05027-20 1 kit (4,000 cm2) 25,000 Blocking One One Chemi-Lumi 䊑䊨䉾䊁䉞䊮䉫ᴺ䉇ELISAᴺ䈭䈬䈱᛫䉕↪䈇䈢∉ቇ⊛ᬌᴺ䈮䈍䈇䈩䇮⋡⊛䉺䊮䊌䉪⾰એᄖ䈱䉺䊮䊌䉪⾰╬䈱㕖․⇣⊛ 䈭ๆ⌕䉕ᛥ䈜䉎䈢䉄䈮䇮䊑䊨䉾䉨䊮䉫ᠲ䈲ᔅⷐਇนᰳ䈪䈅䉍䉁䈜䇯ᧄຠ䈲㜞ಽሶൻว‛䊶䉡䉲↱᧪䉺䊮䊌䉪⾰䉕ේᢱ 䈫䈚䈢䊑䊨䉾䉨䊮䉫ṁᶧ䈪䈅䉍䇮ᓥ᧪䈱䊑䊨䉾䉨䊮䉫ṁᶧ䈫Ყ䈼䇮ఝ䉏䈢ലᨐ䉕⊒ើ䈚䉁䈜䇯 䉁䈢䇮䊕䊦䉥䉨䉲䉻䊷䉷(POD) 䉇䉝䊦䉦䊥䊖䉴䊐䉜䉺䊷䉷(ALP)䈱㉂⚛ᵴᕈ䈮ᓇ㗀䉕ਈ䈋䈭䈇㒐⣣䉕↪䈚䈩䈇䉎䈢䉄䇮㐿ኽᓟ䉅಄⬿䈮䈩ሽ䈜䉎䈖䈫䈏 น⢻䈪䈜䇯 ․˴㐳 ٨ἨἿἕỿὅἂỆᙲẴỦ᧓ửࠢٻỆჺ ٨᭗Ў҄܇ӳཋὉỸἉဌஹἑὅἣἁឋửҾ૰ểẲẺૼẲẟἨἿἕỿὅἂ๋෩ ٨PODẆALPߩᣞእࣱỆࢨ᪪ửɨảễẟ᧸Ᏽдỉ̅ဇỂẆࢸݥờϬᔺ̬܍ӧᏡ ↪ 䊑䊨䉾䉨䊮䉫䈱ᔕᤨ㑆Ყセ ᠲ᧦ઙ 䋨㩼䋩 ᛫࿕ቯ䊒䊧䊷䊃䈱ᚑ 㪈㪇㪇 㪏㪇 㪍㪇 Blocking one ᒰ␠ᓥ᧪ຠ A␠ຠ 㪋㪇 㪉㪇 㪇 㸣㩷㩷䊟䉩᛫䊍䊃䉝䊦䊑䊚䊮᛫㩷㩿㫆㫍㪼㫉㫅㫀㪾㪿㫋㪀 ἨἿἕỿὅἂ 㸣㩷㩷ᧄᶧ5Ꮧ㉼ᶧ㩷㩿ਛ䈱ᔕᤨ㑆㪀 ੑᰴᔕ 㪇 㪌㪇 㸣㩷㩷PODᮡ⼂᛫䊍䊃䉝䊦䊑䊚䊮᛫ (1hrᔕ) ᔕᤨ㑆䋨ಽ䋩 ⊒⦡ᔕ 䈱䉫䊤䊐䈲䊙䉟䉪䊨䊒䊧䊷䊃䈮䈍䈔䉎䊑䊨䉾䉨䊮䉫䈱 ᔕᤨ㑆䈫䊑䊨䉾䉨䊮䉫䈱ലᨐ䈱㑐ଥ䉕䈚䈢䉫䊤䊐䈪䈜䇯 ᒰ␠ᓥ᧪ຠ䋨TBS♽䋩ਗ䈶䈮䌁␠ຠ䈫Ყセ䈚䈩⍴ᤨ㑆 䈪䊑䊨䉾䉨䊮䉫䈏ቢੌ䈚䉁䈜䇯 㸣㩷㩷OPD䈮䉋䉍⊒⦡ (20minᔕ) ᔕᱛ ৻ᰴᔕ㩿䊍䊃䉝䊦䊑䊚䊮ᔕ㪀䉕ⴕ䈦䈩䈇䈭䈇 䈢䉄䇮⊒⦡䈚䈢䉌㕖․⇣ๆ⌕䈏䈖䈦䈩䈇䉎 䈖䈫䉕␜䈜䇯 ↪ߩᵈᗧ 䉝䊎䉳䊮䊶䊎䉥䉼䊮ᔕ䉕↪䈚䈢ᬌ♽䈲䇮ᧄᶧ䈱䉡䉲↱᧪䉺䊮䊌䉪⾰䈫Ꮕᔕ䈜䉎น⢻ᕈ䈏䈅䉍䉁䈜䇯䉁䈢ᧄᶧ䈮 䈘䉏䈩䈇䉎䉺䊮䊌䉪⾰䈲䇮♖䉺䊮䊌䉪⾰䉕↪䈚䈩䈇䉁䈜䈏䇮᷹ቯ♽䈱᛫䉇㉂⚛ᮡ⼂᛫䈭䈬䈫Ꮕᔕ䈜䉎น⢻ᕈ䈏 䈅䉍䉁䈜䇯↪䈮వ┙䈦䈩䇮Ꮕᔕ䈱ή䉕䈗⏕䈒䈣䈘䈇䇯 ଔᩰ ຠฬ Blocking One ⷙᩰ ⾂ᴺ ຠ⇟ภ ኈ㊂ ଔᩰ SP ಄⬿ 03953-95 500 ml 6,000 ߏᵈᗧ˴⹜㛎⎇ⓥ↪એᄖߦߪߏ↪ߒߥߢਅߐޕ ̪ଔᩰߦߪᶖ⾌⒢ߪ߹ࠇߡ߅ࠅ߹ߖࠎޕ ⽼عᄁขᛒᐫ㩷 ࠽ࠞࠗ࠹ࠬࠢᩣᑼળ␠ ˴ޥ੩ㇺᏒਛ੩ੑ᧦ㅢὖਣ᧲₹ደ↸ ࠙ࠚࡉࠨࠗ࠻ http://www.nacalai.co.jp ⹜⮎ߪߎߎߦ ଔᩰ⚊ᦼߩߏᾖળ 0 1 2 0 - 4 8 9 - 5 5 2 ຠߦ㑐ߔࠆᛛⴚ⊛ߥߏᾖળ E-mail:[email protected] TEL:075-211-2703 FAX:075-211-2673 0506
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