小鼠垂體腺苷酸環(huán)化酶激活肽(PACAP)檢測(cè)試劑盒英文名稱Mouse pituitary adenylate cyclase activating polypeptide,PACAP ELISA Kit齊一生物長(zhǎng)期經(jīng)營(yíng)ELISA試劑盒,進(jìn)口ELISA試劑盒及抗體(免費(fèi)代測(cè)),分子學(xué)生物試劑盒,金標(biāo)法檢測(cè)試劑盒,比色法檢測(cè)試劑盒,細(xì)胞菌株,培養(yǎng)基,細(xì)胞因子,生化試劑,耗材等生物試劑產(chǎn)品.誠(chéng)信經(jīng)營(yíng)
小鼠垂體腺苷酸環(huán)化酶激活肽(PACAP)檢測(cè)試劑盒操作步驟:
1、標(biāo)準(zhǔn)品的稀釋:在酶標(biāo)包被板上設(shè)標(biāo)準(zhǔn)品孔10孔,在*、第二孔中分別加標(biāo)準(zhǔn)品100μl,然后在*、第二孔中加標(biāo)準(zhǔn)品稀釋液50μl,混勻;然后從*孔、第二孔中各取100μl分別加到第三孔和第四孔,再在第三、第四孔分別加標(biāo)準(zhǔn)品稀釋液50μl,混勻;然后在第三孔和第四孔中先各取50μl棄掉,再各取50μl分別加到第五、第六孔中,再在第五、第六孔中分別加標(biāo)準(zhǔn)品稀釋液50ul,混勻;混勻后從第五、第六孔中各取50μl分別加到第七、第八孔中,再在第七、第八孔中分別加標(biāo)準(zhǔn)品稀釋液50μl,混勻后從第七、第八孔中分別取50μl加到第九、第十孔中,再在第九第十孔分別加標(biāo)準(zhǔn)品稀釋液50μl,混勻后從第九第十孔中各取50μl棄掉。(稀釋后各孔加樣量都為50μl,濃度分別為9mmol/L,6 mmol/L,3 mmol/L,1.5mmol/L, 0.75 mmol/L)。
2、加樣:分別設(shè)空白孔(空白對(duì)照孔不加樣品及酶標(biāo)試劑,其余各步操作相同)、待測(cè)樣品孔。在酶標(biāo)包被板上待測(cè)樣品孔中先加樣品稀釋液40μl,然后再加待測(cè)樣品10μl(樣品zui終稀釋度為5倍)。加樣將樣品加于酶標(biāo)板孔底部,盡量不觸及孔壁,輕輕晃動(dòng)混勻。
3、溫育:用封板膜封板后置37℃溫育30分鐘。
4、配液:將30(48T的20倍)倍濃縮洗滌液用蒸餾水30(48T的20倍)倍稀釋后備用。
5、洗滌:小心揭掉封板膜,棄去液體,甩干,每孔加滿洗滌液,靜置30秒后棄去,如此重復(fù)5次,拍干。
6、加酶:每孔加入酶標(biāo)試劑50μl,空白孔除外。
7、溫育:操作同3。
8、洗滌:操作同5。
9、顯色:每孔先加入顯色劑A50μl,再加入顯色劑B50μl,輕輕震蕩混勻,37℃避光顯色15分鐘.
10、終止:每孔加終止液50μl,終止反應(yīng)(此時(shí)藍(lán)色立轉(zhuǎn)黃色)。
11、測(cè)定:以空白空調(diào)零,450nm波長(zhǎng)依序測(cè)量各孔的吸光度(OD值)。 測(cè)定應(yīng)在加終止液后15分鐘以內(nèi)進(jìn)行。
操作步驟:
1、標(biāo)準(zhǔn)品的稀釋:在酶標(biāo)包被板上設(shè)標(biāo)準(zhǔn)品孔10孔,在*、第二孔中分別加標(biāo)準(zhǔn)品100μl,然后在*、第二孔中加標(biāo)準(zhǔn)品稀釋液50μl,混勻;然后從*孔、第二孔中各取100μl分別加到第三孔和第四孔,再在第三、第四孔分別加標(biāo)準(zhǔn)品稀釋液50μl,混勻;然后在第三孔和第四孔中先各取50μl棄掉,再各取50μl分別加到第五、第六孔中,再在第五、第六孔中分別加標(biāo)準(zhǔn)品稀釋液50ul,混勻;混勻后從第五、第六孔中各取50μl分別加到第七、第八孔中,再在第七、第八孔中分別加標(biāo)準(zhǔn)品稀釋液50μl,混勻后從第七、第八孔中分別取50μl加到第九、第十孔中,再在第九第十孔分別加標(biāo)準(zhǔn)品稀釋液50μl,混勻后從第九第十孔中各取50μl棄掉。(稀釋后各孔加樣量都為50μl,濃度分別為9mmol/L,6 mmol/L,3 mmol/L,1.5mmol/L, 0.75 mmol/L)。
2、加樣:分別設(shè)空白孔(空白對(duì)照孔不加樣品及酶標(biāo)試劑,其余各步操作相同)、待測(cè)樣品孔。在酶標(biāo)包被板上待測(cè)樣品孔中先加樣品稀釋液40μl,然后再加待測(cè)樣品10μl(樣品zui終稀釋度為5倍)。加樣將樣品加于酶標(biāo)板孔底部,盡量不觸及孔壁,輕輕晃動(dòng)混勻。
3、溫育:用封板膜封板后置37℃溫育30分鐘。
4、配液:將30(48T的20倍)倍濃縮洗滌液用蒸餾水30(48T的20倍)倍稀釋后備用。
5、洗滌:小心揭掉封板膜,棄去液體,甩干,每孔加滿洗滌液,靜置30秒后棄去,如此重復(fù)5次,拍干。
6、加酶:每孔加入酶標(biāo)試劑50μl,空白孔除外。
7、溫育:操作同3。
8、洗滌:操作同5。
9、顯色:每孔先加入顯色劑A50μl,再加入顯色劑B50μl,輕輕震蕩混勻,37℃避光顯色15分鐘.
10、終止:每孔加終止液50μl,終止反應(yīng)(此時(shí)藍(lán)色立轉(zhuǎn)黃色)。
11、測(cè)定:以空白空調(diào)零,450nm波長(zhǎng)依序測(cè)量各孔的吸光度(OD值)。 測(cè)定應(yīng)在加終止液后15分鐘以內(nèi)進(jìn)行。15082 Microtube Racked Systems 10 RACKS/PKG
15086 Microtubes (not racked) 960/PKG
15088 Microtube 8-Cap Strips 120/PKG
15100 Amine-binding, Maleic Anhydride Activated Plates (clear, 8-well strips) PKG OF 5
15102 Amine-binding, Maleic Anhydride Activated Plates (clear, 8-well strips), 5 × 5 ea 5 X 5 EACH
15108 Amine-binding, Maleic Anhydride Activated White 96-Well Plates 5 PLATES
15110 Amine-binding, Maleic Anhydride Activated Plates (clear, 96-well) PKG OF 5
15112 Amine-binding, Maleic Anhydride Activated Plates (clear, 96-well), 5 × 5 ea 5 X 5 EACH
15116 NeutrAvidin Coated Plates, 96-well, white 5 PLATES
15117 NeutrAvidin Coated Plates, 96-well, black 5 PLATES
15118 Streptavidin Coated Plates, 96-well, white 5 PLATES
15119 Streptavidin Coated Plates, 96-well, black 5 PLATES
15120 Streptavidin Coated Plates, 96-well strip, clear PKG OF 5
15121 Streptavidin Coated Plates, 96-well strip, clear 5 PLATES
15122 Streptavidin Coated Plates, 96-well strip, clear 25/PKG (5X5)
15123 NeutrAvidin Coated Plates, 96-well, clear 5 PLATES
15124 Streptavidin Coated Plates, 96-well, clear PKG OF 5
15125 Streptavidin Coated Plates, 96-well, clear 5 PLATES
15126 Streptavidin Coated Plates, 96-well, clear PKG OF 25
15128 NeutrAvidin Coated Plates, 96-well strip, clear 5 PLATES/EA
15130 Protein A Coated Clear 96-Well Plates 5 PLATES
15131 Protein G Coated Clear 96-Well Plates 5 PLATES
15132 Protein A Coated Strip Plates 5 PLATES
15133 Protein G Coated Strip Plates 5 PLATES
15134 Goat Anti-Mouse IgG Coated Clear 96-well Plates 5 PLATES
15135 Goat Anti-Rabbit IgG Coated Plates, 96-well, clear 5 PLATES
15136 Goat Anti-Rabbit IgG Coated Plates, 96-well, white 5 PLATES
15137 Goat Anti-Rabbit IgG Coated Plates, 96-well, black 5 PLATES
15138 Protein A/G Coated Plates 5 PLATES
15140 Pierce Glutathione Coated Strip Plates, (8-well strips) 5 PLATES
15142 Nickel Coated Plates, 96-Well Strip, Clear 5 PLATES
15143 Copper Coated, High Binding Capacity Plates (clear, 96-well) 5 PLATES
15144 Anti-GST Coated Plates, Clear, 8-Well Strip 5 plates
15146 Copper Coated, High Binding Capacity Plates (clear, 8-well strips) 5 PLT, STRIP
15147 Copper Coated, High Binding Capacity Plates (white, 96-well) 5 PLATES
15148 Copper Coated, High Binding Capacity Plates (black, 96-well) 5 PLATES
15150 Maleimide Activated Plates, Clear 8-well Strip Plates 5 PLATES
15151 Biotin Coated Plates, 96-well strip 5 PLATES
15152 Maleimide Activated White 96-Well Plates 5 PLATES
15153 Maleimide Activated Black 96-Well Plates 5 PLATES
15154 Protein A Coated White 96-Well Plates 5 PLATES
15155 Protein A Coated Black 96-Well Plates 5 PLATES
15156 Protein G Coated White 96-Well Plates 5 PLATES
15157 Protein G Coated Black 96-Well Plates 5 PLATES
15159 QuantaRed Enhanced Chemifluorescent HRP Substrate Kit 110mL kit
15162 QuantaBlu NS/K Substrate (for nonstopped and kinetic assays) Kit
15165 HisProbe-HRP Conjμgate 2 mg
15168 SuperSignal West HisProbe Kit 685mL kit
15169 QuantaBlu Fluorogenic Peroxidase Substrate Kit Kit
15190 Protein L Coated Clear 96-Well Plates 5 PLATES
15216 NeutrAvidin Coated Plates, 96-well, white 5 PLATES
15217 NeutrAvidin Coated Plates, 96-well, black 5 PLATES
15218 Streptavidin Coated Plates, 96-well, white 5 PLATES
15219 Streptavidin Coated Plates, 96-well, black 5 PLATES
15234 Goat Anti-Mouse IgG Coated White 96-well Plates 5 PLATES
15240 Pierce Glutathione Coated Plates, (white, 96-well) 5 PLATES
15242 Nickel Coated Plates, 96-Well, White 5 PLATES