猴腫瘤標(biao)(biao)(biao)(biao)志物(wu)(wu)(wu)CA724(CA724)酶(mei)(mei)聯免疫試(shi)劑(ji)盒 實驗(yan)(yan)原(yuan)理(li)(li) ELISA運用(yong)了免疫學原(yuan)理(li)(li)和(he)(he)化學反應(ying)顯色原(yuan)理(li)(li),需(xu)要將抗(kang)原(yuan)或抗(kang)體(ti)預包(bao)被在(zai)(zai)(zai)(zai)(zai)固相載(zai)體(ti)表面,使后(hou)來形成的(de)(de)抗(kang)原(yuan)-抗(kang)體(ti)-酶(mei)(mei) (熒光(guang)基團) -底(di)物(wu)(wu)(wu)復(fu)合物(wu)(wu)(wu)黏附在(zai)(zai)(zai)(zai)(zai)載(zai)體(ti)上,通過檢測(ce)(ce)(ce)OD值(zhi)或發光(guang)值(zhi),來檢測(ce)(ce)(ce)靶蛋白(bai)的(de)(de)含量。 實驗(yan)(yan)所需(xu)自備物(wu)(wu)(wu)品(pin)1. 酶(mei)(mei)標(biao)(biao)(biao)(biao)儀(450nm)2. 加(jia)樣(yang)(yang)器(qi)及(ji)槍頭:0.5-10uL、2-20uL、20-200uL、200-1000uL3. 37℃恒溫(wen)(wen)(wen)(wen)箱 試(shi)劑(ji)盒組成名稱96 孔配(pei)置(zhi)48 孔配(pei)置(zhi)備注(zhu)微孔酶(mei)(mei)標(biao)(biao)(biao)(biao)板(ban)(ban)12 孔×8 條12 孔×4 條無(wu)標(biao)(biao)(biao)(biao)準(zhun)品(pin)0.5mL0.5mL按說(shuo)(shuo)明(ming)書(shu)進(jin)(jin)行稀(xi)釋(shi)標(biao)(biao)(biao)(biao)準(zhun)品(pin)稀(xi)釋(shi)液(ye)(ye)(ye)6mL3mL按說(shuo)(shuo)明(ming)書(shu)進(jin)(jin)行稀(xi)釋(shi)樣(yang)(yang)本(ben)(ben)稀(xi)釋(shi)液(ye)(ye)(ye)6mL3mL按說(shuo)(shuo)明(ming)書(shu)進(jin)(jin)行稀(xi)釋(shi)檢測(ce)(ce)(ce)抗(kang)體(ti)-HRP6mL3mL無(wu)20×洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)緩(huan)沖液(ye)(ye)(ye)20mL20mL按說(shuo)(shuo)明(ming)書(shu)進(jin)(jin)行稀(xi)釋(shi)底(di)物(wu)(wu)(wu) A6mL3mL無(wu)底(di)物(wu)(wu)(wu) B6mL3mL無(wu)終(zhong)(zhong)止液(ye)(ye)(ye)6mL3mL無(wu)封板(ban)(ban)膜(mo)2 張2 張無(wu)說(shuo)(shuo)明(ming)書(shu)1 份1 份無(wu)自封袋1 個(ge)(ge)(ge)(ge)1 個(ge)(ge)(ge)(ge)無(wu) 操(cao)作(zuo)步驟1. 準(zhun)備工作(zuo):將試(shi)劑(ji)盒從(cong)冰(bing)箱中(zhong)(zhong)取出(chu)(chu),室(shi)溫(wen)(wen)(wen)(wen)放置(zhi)30分(fen)(fen)鐘,使試(shi)劑(ji)盒恢復(fu)至室(shi)溫(wen)(wen)(wen)(wen)。同時準(zhun)備好實驗(yan)(yan)所需(xu)的(de)(de)材料(liao)和(he)(he)器(qi)具。2. 加(jia)樣(yang)(yang):在(zai)(zai)(zai)(zai)(zai)酶(mei)(mei)標(biao)(biao)(biao)(biao)板(ban)(ban)的(de)(de)弟1個(ge)(ge)(ge)(ge)孔中(zhong)(zhong)加(jia)入標(biao)(biao)(biao)(biao)準(zhun)品(pin)50μL,然后(hou)在(zai)(zai)(zai)(zai)(zai)后(hou)續的(de)(de)孔中(zhong)(zhong)依(yi)次(ci)(ci)加(jia)入待測(ce)(ce)(ce)樣(yang)(yang)本(ben)(ben)50μL。3. 溫(wen)(wen)(wen)(wen)育:將酶(mei)(mei)標(biao)(biao)(biao)(biao)板(ban)(ban)密封,在(zai)(zai)(zai)(zai)(zai)37℃下(xia)溫(wen)(wen)(wen)(wen)育30分(fen)(fen)鐘。4. 洗(xi)(xi)(xi)(xi)(xi)滌(di)(di):用(yong)洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)液(ye)(ye)(ye)清(qing)洗(xi)(xi)(xi)(xi)(xi)酶(mei)(mei)標(biao)(biao)(biao)(biao)板(ban)(ban),共(gong)洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)5次(ci)(ci)。每(mei)次(ci)(ci)洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)后(hou),用(yong)吸(xi)水紙輕(qing)(qing)輕(qing)(qing)吸(xi)干孔內液(ye)(ye)(ye)體(ti)。5. 加(jia)酶(mei)(mei):在(zai)(zai)(zai)(zai)(zai)每(mei)個(ge)(ge)(ge)(ge)孔中(zhong)(zhong)加(jia)入酶(mei)(mei)標(biao)(biao)(biao)(biao)物(wu)(wu)(wu)100μL。6. 溫(wen)(wen)(wen)(wen)育:再次(ci)(ci)將酶(mei)(mei)標(biao)(biao)(biao)(biao)板(ban)(ban)密封,在(zai)(zai)(zai)(zai)(zai)37℃下(xia)溫(wen)(wen)(wen)(wen)育30分(fen)(fen)鐘。7. 洗(xi)(xi)(xi)(xi)(xi)滌(di)(di):用(yong)洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)液(ye)(ye)(ye)清(qing)洗(xi)(xi)(xi)(xi)(xi)酶(mei)(mei)標(biao)(biao)(biao)(biao)板(ban)(ban),共(gong)洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)5次(ci)(ci)。每(mei)次(ci)(ci)洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)后(hou),用(yong)吸(xi)水紙輕(qing)(qing)輕(qing)(qing)吸(xi)干孔內液(ye)(ye)(ye)體(ti)。8. 加(jia)底(di)物(wu)(wu)(wu):在(zai)(zai)(zai)(zai)(zai)每(mei)個(ge)(ge)(ge)(ge)孔中(zhong)(zhong)加(jia)入底(di)物(wu)(wu)(wu)A液(ye)(ye)(ye)50μL,再加(jia)入底(di)物(wu)(wu)(wu)B液(ye)(ye)(ye)50μL。9. 終(zhong)(zhong)止反應(ying):在(zai)(zai)(zai)(zai)(zai)每(mei)個(ge)(ge)(ge)(ge)孔中(zhong)(zhong)加(jia)入終(zhong)(zhong)止液(ye)(ye)(ye)50μL。10. 檢測(ce)(ce)(ce):用(yong)酶(mei)(mei)標(biao)(biao)(biao)(biao)儀在(zai)(zai)(zai)(zai)(zai)450nm波長下(xia)讀取各孔的(de)(de)吸(xi)光(guang)度(OD值(zhi))。 結果分(fen)(fen)析根據(ju)標(biao)(biao)(biao)(biao)準(zhun)品(pin)和(he)(he)樣(yang)(yang)本(ben)(ben)的(de)(de)OD值(zhi),繪制標(biao)(biao)(biao)(biao)準(zhun)曲線,從(cong)而計算出(chu)(chu)樣(yang)(yang)本(ben)(ben)中(zhong)(zhong)角蛋白(bai)1(KRT1)的(de)(de)濃度。具體(ti)分(fen)(fen)析方(fang)法可參考試(shi)劑(ji)盒說(shuo)(shuo)明(ming)書(shu)或相關(guan)文(wen)獻(xian)。注(zhu)意(yi)事項(xiang)1. 在(zai)(zai)(zai)(zai)(zai)操(cao)作(zuo)過程(cheng)中(zhong)(zhong),應(ying)保(bao)持酶(mei)(mei)標(biao)(biao)(biao)(biao)板(ban)(ban)的(de)(de)干燥,避免水分(fen)(fen)進(jin)(jin)入孔內。2. 加(jia)樣(yang)(yang)時需(xu)確(que)(que)保(bao)加(jia)樣(yang)(yang)準(zhun)確(que)(que)、無(wu)氣泡(pao)產(chan)(chan)生(sheng)(sheng)。3. 溫(wen)(wen)(wen)(wen)育和(he)(he)洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)過程(cheng)中(zhong)(zhong)需(xu)嚴格(ge)控制時間和(he)(he)溫(wen)(wen)(wen)(wen)度,確(que)(que)保(bao)實驗(yan)(yan)結果的(de)(de)準(zhun)確(que)(que)性(xing)。4. 在(zai)(zai)(zai)(zai)(zai)使用(yong)酶(mei)(mei)標(biao)(biao)(biao)(biao)儀讀取OD值(zhi)時,需(xu)確(que)(que)保(bao)波長設置(zhi)正確(que)(que),避免誤差產(chan)(chan)生(sheng)(sheng)。5. 在(zai)(zai)(zai)(zai)(zai)整個(ge)(ge)(ge)(ge)實驗(yan)(yan)過程(cheng)中(zhong)(zhong),需(xu)保(bao)持操(cao)作(zuo)環(huan)境的(de)(de)清(qing)潔和(he)(he)整潔,避免交叉污染。
猴胎兒血紅(hong)蛋(dan)白(bai)(HBF)酶(mei)(mei)聯免(mian)(mian)(mian)(mian)疫試劑(ji)盒(he) 實(shi)驗(yan)原(yuan)(yuan)理 ELISA運(yun)用(yong)(yong)(yong)了免(mian)(mian)(mian)(mian)疫學原(yuan)(yuan)理和(he)(he)化學反(fan)應(ying)顯(xian)色原(yuan)(yuan)理,需(xu)要(yao)將抗(kang)原(yuan)(yuan)或(huo)抗(kang)體(ti)(ti)(ti)預包被在(zai)(zai)(zai)固相載體(ti)(ti)(ti)表(biao)面,使(shi)后來形(xing)成(cheng)的(de)(de)(de)抗(kang)原(yuan)(yuan)-抗(kang)體(ti)(ti)(ti)-酶(mei)(mei) (熒光基團) -底物(wu)(wu)(wu)復合物(wu)(wu)(wu)黏附在(zai)(zai)(zai)載體(ti)(ti)(ti)上,通過檢測(ce)(ce)OD值或(huo)發光值,來檢測(ce)(ce)靶蛋(dan)白(bai)的(de)(de)(de)含量。 實(shi)驗(yan)所(suo)需(xu)自(zi)備(bei)物(wu)(wu)(wu)品1. 酶(mei)(mei)標(biao)(biao)(biao)(biao)儀(450nm)2. 加(jia)樣(yang)(yang)器(qi)及槍頭:0.5-10uL、2-20uL、20-200uL、200-1000uL3. 37℃恒溫箱 試劑(ji)盒(he)組成(cheng)名稱96 孔(kong)(kong)(kong)(kong)配置(zhi)(zhi)48 孔(kong)(kong)(kong)(kong)配置(zhi)(zhi)備(bei)注微孔(kong)(kong)(kong)(kong)酶(mei)(mei)標(biao)(biao)(biao)(biao)板(ban)(ban)12 孔(kong)(kong)(kong)(kong)×8 條(tiao)12 孔(kong)(kong)(kong)(kong)×4 條(tiao)無(wu)(wu)(wu)標(biao)(biao)(biao)(biao)準(zhun)品0.5mL0.5mL按(an)說(shuo)明書進(jin)行(xing)(xing)稀(xi)釋(shi)(shi)(shi)標(biao)(biao)(biao)(biao)準(zhun)品稀(xi)釋(shi)(shi)(shi)液(ye)6mL3mL按(an)說(shuo)明書進(jin)行(xing)(xing)稀(xi)釋(shi)(shi)(shi)樣(yang)(yang)本(ben)稀(xi)釋(shi)(shi)(shi)液(ye)6mL3mL按(an)說(shuo)明書進(jin)行(xing)(xing)稀(xi)釋(shi)(shi)(shi)檢測(ce)(ce)抗(kang)體(ti)(ti)(ti)-HRP6mL3mL無(wu)(wu)(wu)20×洗(xi)(xi)滌(di)(di)(di)緩沖液(ye)20mL20mL按(an)說(shuo)明書進(jin)行(xing)(xing)稀(xi)釋(shi)(shi)(shi)底物(wu)(wu)(wu) A6mL3mL無(wu)(wu)(wu)底物(wu)(wu)(wu) B6mL3mL無(wu)(wu)(wu)終(zhong)止液(ye)6mL3mL無(wu)(wu)(wu)封板(ban)(ban)膜(mo)2 張2 張無(wu)(wu)(wu)說(shuo)明書1 份1 份無(wu)(wu)(wu)自(zi)封袋1 個(ge)1 個(ge)無(wu)(wu)(wu) 操(cao)(cao)作(zuo)步驟(zou)1. 準(zhun)備(bei)工(gong)作(zuo):將試劑(ji)盒(he)從(cong)冰箱中(zhong)(zhong)(zhong)(zhong)取出(chu),室(shi)溫放置(zhi)(zhi)30分鐘,使(shi)試劑(ji)盒(he)恢(hui)復至室(shi)溫。同(tong)時準(zhun)備(bei)好(hao)實(shi)驗(yan)所(suo)需(xu)的(de)(de)(de)材料和(he)(he)器(qi)具。2. 加(jia)樣(yang)(yang):在(zai)(zai)(zai)酶(mei)(mei)標(biao)(biao)(biao)(biao)板(ban)(ban)的(de)(de)(de)弟(di)1個(ge)孔(kong)(kong)(kong)(kong)中(zhong)(zhong)(zhong)(zhong)加(jia)入(ru)(ru)標(biao)(biao)(biao)(biao)準(zhun)品50μL,然后在(zai)(zai)(zai)后續的(de)(de)(de)孔(kong)(kong)(kong)(kong)中(zhong)(zhong)(zhong)(zhong)依(yi)次(ci)加(jia)入(ru)(ru)待測(ce)(ce)樣(yang)(yang)本(ben)50μL。3. 溫育:將酶(mei)(mei)標(biao)(biao)(biao)(biao)板(ban)(ban)密封,在(zai)(zai)(zai)37℃下(xia)溫育30分鐘。4. 洗(xi)(xi)滌(di)(di)(di):用(yong)(yong)(yong)洗(xi)(xi)滌(di)(di)(di)液(ye)清洗(xi)(xi)酶(mei)(mei)標(biao)(biao)(biao)(biao)板(ban)(ban),共(gong)洗(xi)(xi)滌(di)(di)(di)5次(ci)。每(mei)(mei)次(ci)洗(xi)(xi)滌(di)(di)(di)后,用(yong)(yong)(yong)吸(xi)水紙(zhi)(zhi)輕(qing)(qing)輕(qing)(qing)吸(xi)干(gan)孔(kong)(kong)(kong)(kong)內(nei)(nei)液(ye)體(ti)(ti)(ti)。5. 加(jia)酶(mei)(mei):在(zai)(zai)(zai)每(mei)(mei)個(ge)孔(kong)(kong)(kong)(kong)中(zhong)(zhong)(zhong)(zhong)加(jia)入(ru)(ru)酶(mei)(mei)標(biao)(biao)(biao)(biao)物(wu)(wu)(wu)100μL。6. 溫育:再次(ci)將酶(mei)(mei)標(biao)(biao)(biao)(biao)板(ban)(ban)密封,在(zai)(zai)(zai)37℃下(xia)溫育30分鐘。7. 洗(xi)(xi)滌(di)(di)(di):用(yong)(yong)(yong)洗(xi)(xi)滌(di)(di)(di)液(ye)清洗(xi)(xi)酶(mei)(mei)標(biao)(biao)(biao)(biao)板(ban)(ban),共(gong)洗(xi)(xi)滌(di)(di)(di)5次(ci)。每(mei)(mei)次(ci)洗(xi)(xi)滌(di)(di)(di)后,用(yong)(yong)(yong)吸(xi)水紙(zhi)(zhi)輕(qing)(qing)輕(qing)(qing)吸(xi)干(gan)孔(kong)(kong)(kong)(kong)內(nei)(nei)液(ye)體(ti)(ti)(ti)。8. 加(jia)底物(wu)(wu)(wu):在(zai)(zai)(zai)每(mei)(mei)個(ge)孔(kong)(kong)(kong)(kong)中(zhong)(zhong)(zhong)(zhong)加(jia)入(ru)(ru)底物(wu)(wu)(wu)A液(ye)50μL,再加(jia)入(ru)(ru)底物(wu)(wu)(wu)B液(ye)50μL。9. 終(zhong)止反(fan)應(ying):在(zai)(zai)(zai)每(mei)(mei)個(ge)孔(kong)(kong)(kong)(kong)中(zhong)(zhong)(zhong)(zhong)加(jia)入(ru)(ru)終(zhong)止液(ye)50μL。10. 檢測(ce)(ce):用(yong)(yong)(yong)酶(mei)(mei)標(biao)(biao)(biao)(biao)儀在(zai)(zai)(zai)450nm波(bo)長下(xia)讀取各(ge)孔(kong)(kong)(kong)(kong)的(de)(de)(de)吸(xi)光度(OD值)。 結果(guo)分析根據(ju)標(biao)(biao)(biao)(biao)準(zhun)品和(he)(he)樣(yang)(yang)本(ben)的(de)(de)(de)OD值,繪制標(biao)(biao)(biao)(biao)準(zhun)曲(qu)線,從(cong)而計(ji)算出(chu)樣(yang)(yang)本(ben)中(zhong)(zhong)(zhong)(zhong)角(jiao)蛋(dan)白(bai)1(KRT1)的(de)(de)(de)濃(nong)度。具體(ti)(ti)(ti)分析方(fang)法(fa)可(ke)參考試劑(ji)盒(he)說(shuo)明書或(huo)相關(guan)文獻。注意事項1. 在(zai)(zai)(zai)操(cao)(cao)作(zuo)過程(cheng)中(zhong)(zhong)(zhong)(zhong),應(ying)保(bao)持酶(mei)(mei)標(biao)(biao)(biao)(biao)板(ban)(ban)的(de)(de)(de)干(gan)燥(zao),避(bi)免(mian)(mian)(mian)(mian)水分進(jin)入(ru)(ru)孔(kong)(kong)(kong)(kong)內(nei)(nei)。2. 加(jia)樣(yang)(yang)時需(xu)確(que)保(bao)加(jia)樣(yang)(yang)準(zhun)確(que)、無(wu)(wu)(wu)氣泡(pao)產生。3. 溫育和(he)(he)洗(xi)(xi)滌(di)(di)(di)過程(cheng)中(zhong)(zhong)(zhong)(zhong)需(xu)嚴格控制時間和(he)(he)溫度,確(que)保(bao)實(shi)驗(yan)結果(guo)的(de)(de)(de)準(zhun)確(que)性。4. 在(zai)(zai)(zai)使(shi)用(yong)(yong)(yong)酶(mei)(mei)標(biao)(biao)(biao)(biao)儀讀取OD值時,需(xu)確(que)保(bao)波(bo)長設置(zhi)(zhi)正確(que),避(bi)免(mian)(mian)(mian)(mian)誤差產生。5. 在(zai)(zai)(zai)整個(ge)實(shi)驗(yan)過程(cheng)中(zhong)(zhong)(zhong)(zhong),需(xu)保(bao)持操(cao)(cao)作(zuo)環境的(de)(de)(de)清潔(jie)和(he)(he)整潔(jie),避(bi)免(mian)(mian)(mian)(mian)交叉污染。
猴(hou)原(yuan)(yuan)纖維蛋白2(FBN2)酶(mei)(mei)(mei)聯免(mian)疫(yi)試(shi)劑(ji)盒(he)實驗(yan)(yan)原(yuan)(yuan)理(li) ELISA運(yun)用(yong)了免(mian)疫(yi)學原(yuan)(yuan)理(li)和化學反應(ying)顯(xian)色原(yuan)(yuan)理(li),需(xu)要將(jiang)抗(kang)(kang)原(yuan)(yuan)或(huo)抗(kang)(kang)體(ti)(ti)預包被在(zai)(zai)(zai)(zai)(zai)固(gu)相(xiang)載體(ti)(ti)表(biao)面(mian),使(shi)(shi)后(hou)來形成的(de)(de)抗(kang)(kang)原(yuan)(yuan)-抗(kang)(kang)體(ti)(ti)-酶(mei)(mei)(mei) (熒光(guang)基團) -底(di)物(wu)復合(he)物(wu)黏(nian)附在(zai)(zai)(zai)(zai)(zai)載體(ti)(ti)上,通過(guo)檢(jian)(jian)測OD值(zhi)(zhi)或(huo)發光(guang)值(zhi)(zhi),來檢(jian)(jian)測靶蛋白的(de)(de)含(han)量。 實驗(yan)(yan)所需(xu)自(zi)備(bei)物(wu)品(pin)1. 酶(mei)(mei)(mei)標(biao)(biao)(biao)儀(450nm)2. 加(jia)(jia)樣(yang)器及槍頭:0.5-10uL、2-20uL、20-200uL、200-1000uL3. 37℃恒溫(wen)(wen)(wen)(wen)箱 試(shi)劑(ji)盒(he)組(zu)成名稱96 孔(kong)(kong)(kong)配置(zhi)48 孔(kong)(kong)(kong)配置(zhi)備(bei)注(zhu)(zhu)微孔(kong)(kong)(kong)酶(mei)(mei)(mei)標(biao)(biao)(biao)板(ban)(ban)12 孔(kong)(kong)(kong)×8 條12 孔(kong)(kong)(kong)×4 條無(wu)(wu)(wu)標(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)0.5mL0.5mL按說(shuo)(shuo)(shuo)明書(shu)(shu)進(jin)行(xing)(xing)稀(xi)釋(shi)(shi)(shi)標(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)稀(xi)釋(shi)(shi)(shi)液6mL3mL按說(shuo)(shuo)(shuo)明書(shu)(shu)進(jin)行(xing)(xing)稀(xi)釋(shi)(shi)(shi)樣(yang)本(ben)稀(xi)釋(shi)(shi)(shi)液6mL3mL按說(shuo)(shuo)(shuo)明書(shu)(shu)進(jin)行(xing)(xing)稀(xi)釋(shi)(shi)(shi)檢(jian)(jian)測抗(kang)(kang)體(ti)(ti)-HRP6mL3mL無(wu)(wu)(wu)20×洗(xi)(xi)(xi)滌(di)(di)(di)緩沖液20mL20mL按說(shuo)(shuo)(shuo)明書(shu)(shu)進(jin)行(xing)(xing)稀(xi)釋(shi)(shi)(shi)底(di)物(wu) A6mL3mL無(wu)(wu)(wu)底(di)物(wu) B6mL3mL無(wu)(wu)(wu)終(zhong)止液6mL3mL無(wu)(wu)(wu)封板(ban)(ban)膜2 張2 張無(wu)(wu)(wu)說(shuo)(shuo)(shuo)明書(shu)(shu)1 份1 份無(wu)(wu)(wu)自(zi)封袋1 個1 個無(wu)(wu)(wu) 操作步驟1. 準(zhun)(zhun)(zhun)備(bei)工作:將(jiang)試(shi)劑(ji)盒(he)從(cong)冰箱中(zhong)取出,室溫(wen)(wen)(wen)(wen)放置(zhi)30分鐘(zhong)(zhong),使(shi)(shi)試(shi)劑(ji)盒(he)恢復至室溫(wen)(wen)(wen)(wen)。同時(shi)準(zhun)(zhun)(zhun)備(bei)好實驗(yan)(yan)所需(xu)的(de)(de)材料(liao)和器具。2. 加(jia)(jia)樣(yang):在(zai)(zai)(zai)(zai)(zai)酶(mei)(mei)(mei)標(biao)(biao)(biao)板(ban)(ban)的(de)(de)弟(di)1個孔(kong)(kong)(kong)中(zhong)加(jia)(jia)入(ru)(ru)標(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)50μL,然后(hou)在(zai)(zai)(zai)(zai)(zai)后(hou)續的(de)(de)孔(kong)(kong)(kong)中(zhong)依次(ci)加(jia)(jia)入(ru)(ru)待測樣(yang)本(ben)50μL。3. 溫(wen)(wen)(wen)(wen)育(yu)(yu):將(jiang)酶(mei)(mei)(mei)標(biao)(biao)(biao)板(ban)(ban)密(mi)封,在(zai)(zai)(zai)(zai)(zai)37℃下溫(wen)(wen)(wen)(wen)育(yu)(yu)30分鐘(zhong)(zhong)。4. 洗(xi)(xi)(xi)滌(di)(di)(di):用(yong)洗(xi)(xi)(xi)滌(di)(di)(di)液清(qing)(qing)洗(xi)(xi)(xi)酶(mei)(mei)(mei)標(biao)(biao)(biao)板(ban)(ban),共洗(xi)(xi)(xi)滌(di)(di)(di)5次(ci)。每(mei)(mei)次(ci)洗(xi)(xi)(xi)滌(di)(di)(di)后(hou),用(yong)吸水紙輕輕吸干孔(kong)(kong)(kong)內液體(ti)(ti)。5. 加(jia)(jia)酶(mei)(mei)(mei):在(zai)(zai)(zai)(zai)(zai)每(mei)(mei)個孔(kong)(kong)(kong)中(zhong)加(jia)(jia)入(ru)(ru)酶(mei)(mei)(mei)標(biao)(biao)(biao)物(wu)100μL。6. 溫(wen)(wen)(wen)(wen)育(yu)(yu):再(zai)次(ci)將(jiang)酶(mei)(mei)(mei)標(biao)(biao)(biao)板(ban)(ban)密(mi)封,在(zai)(zai)(zai)(zai)(zai)37℃下溫(wen)(wen)(wen)(wen)育(yu)(yu)30分鐘(zhong)(zhong)。7. 洗(xi)(xi)(xi)滌(di)(di)(di):用(yong)洗(xi)(xi)(xi)滌(di)(di)(di)液清(qing)(qing)洗(xi)(xi)(xi)酶(mei)(mei)(mei)標(biao)(biao)(biao)板(ban)(ban),共洗(xi)(xi)(xi)滌(di)(di)(di)5次(ci)。每(mei)(mei)次(ci)洗(xi)(xi)(xi)滌(di)(di)(di)后(hou),用(yong)吸水紙輕輕吸干孔(kong)(kong)(kong)內液體(ti)(ti)。8. 加(jia)(jia)底(di)物(wu):在(zai)(zai)(zai)(zai)(zai)每(mei)(mei)個孔(kong)(kong)(kong)中(zhong)加(jia)(jia)入(ru)(ru)底(di)物(wu)A液50μL,再(zai)加(jia)(jia)入(ru)(ru)底(di)物(wu)B液50μL。9. 終(zhong)止反應(ying):在(zai)(zai)(zai)(zai)(zai)每(mei)(mei)個孔(kong)(kong)(kong)中(zhong)加(jia)(jia)入(ru)(ru)終(zhong)止液50μL。10. 檢(jian)(jian)測:用(yong)酶(mei)(mei)(mei)標(biao)(biao)(biao)儀在(zai)(zai)(zai)(zai)(zai)450nm波長下讀取各孔(kong)(kong)(kong)的(de)(de)吸光(guang)度(du)(OD值(zhi)(zhi))。 結果(guo)分析根據標(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)和樣(yang)本(ben)的(de)(de)OD值(zhi)(zhi),繪制標(biao)(biao)(biao)準(zhun)(zhun)(zhun)曲線,從(cong)而(er)計(ji)算(suan)出樣(yang)本(ben)中(zhong)角蛋白1(KRT1)的(de)(de)濃度(du)。具體(ti)(ti)分析方法可參考試(shi)劑(ji)盒(he)說(shuo)(shuo)(shuo)明書(shu)(shu)或(huo)相(xiang)關(guan)文獻。注(zhu)(zhu)意事(shi)項1. 在(zai)(zai)(zai)(zai)(zai)操作過(guo)程中(zhong),應(ying)保(bao)(bao)持(chi)酶(mei)(mei)(mei)標(biao)(biao)(biao)板(ban)(ban)的(de)(de)干燥(zao),避(bi)免(mian)水分進(jin)入(ru)(ru)孔(kong)(kong)(kong)內。2. 加(jia)(jia)樣(yang)時(shi)需(xu)確(que)保(bao)(bao)加(jia)(jia)樣(yang)準(zhun)(zhun)(zhun)確(que)、無(wu)(wu)(wu)氣泡產生。3. 溫(wen)(wen)(wen)(wen)育(yu)(yu)和洗(xi)(xi)(xi)滌(di)(di)(di)過(guo)程中(zhong)需(xu)嚴格控制時(shi)間和溫(wen)(wen)(wen)(wen)度(du),確(que)保(bao)(bao)實驗(yan)(yan)結果(guo)的(de)(de)準(zhun)(zhun)(zhun)確(que)性。4. 在(zai)(zai)(zai)(zai)(zai)使(shi)(shi)用(yong)酶(mei)(mei)(mei)標(biao)(biao)(biao)儀讀取OD值(zhi)(zhi)時(shi),需(xu)確(que)保(bao)(bao)波長設(she)置(zhi)正確(que),避(bi)免(mian)誤(wu)差(cha)產生。5. 在(zai)(zai)(zai)(zai)(zai)整個實驗(yan)(yan)過(guo)程中(zhong),需(xu)保(bao)(bao)持(chi)操作環(huan)境的(de)(de)清(qing)(qing)潔和整潔,避(bi)免(mian)交(jiao)叉(cha)污染(ran)。
猴糖皮質激(ji)素受體(ti)α(GRα)酶(mei)(mei)(mei)(mei)聯免疫試(shi)劑盒(he) 實(shi)(shi)驗(yan)原(yuan)理(li)(li) ELISA運用(yong)(yong)了免疫學(xue)原(yuan)理(li)(li)和(he)(he)化學(xue)反應顯色原(yuan)理(li)(li),需(xu)(xu)要將(jiang)抗(kang)(kang)原(yuan)或抗(kang)(kang)體(ti)預包被在(zai)固相載(zai)體(ti)表(biao)面,使后來形成(cheng)的(de)(de)(de)抗(kang)(kang)原(yuan)-抗(kang)(kang)體(ti)-酶(mei)(mei)(mei)(mei) (熒光(guang)基(ji)團) -底(di)物(wu)(wu)(wu)復(fu)合物(wu)(wu)(wu)黏附在(zai)載(zai)體(ti)上,通(tong)過檢(jian)測OD值或發光(guang)值,來檢(jian)測靶蛋(dan)白(bai)的(de)(de)(de)含量。 實(shi)(shi)驗(yan)所(suo)需(xu)(xu)自備(bei)(bei)物(wu)(wu)(wu)品(pin)(pin)1. 酶(mei)(mei)(mei)(mei)標(biao)儀(450nm)2. 加(jia)(jia)(jia)(jia)(jia)樣(yang)(yang)器及槍頭:0.5-10uL、2-20uL、20-200uL、200-1000uL3. 37℃恒溫(wen)(wen)(wen)箱 試(shi)劑盒(he)組(zu)成(cheng)名稱96 孔(kong)(kong)配置(zhi)(zhi)48 孔(kong)(kong)配置(zhi)(zhi)備(bei)(bei)注微孔(kong)(kong)酶(mei)(mei)(mei)(mei)標(biao)板(ban)(ban)(ban)12 孔(kong)(kong)×8 條12 孔(kong)(kong)×4 條無(wu)標(biao)準品(pin)(pin)0.5mL0.5mL按(an)(an)說(shuo)明(ming)書(shu)(shu)進行(xing)稀釋標(biao)準品(pin)(pin)稀釋液6mL3mL按(an)(an)說(shuo)明(ming)書(shu)(shu)進行(xing)稀釋樣(yang)(yang)本稀釋液6mL3mL按(an)(an)說(shuo)明(ming)書(shu)(shu)進行(xing)稀釋檢(jian)測抗(kang)(kang)體(ti)-HRP6mL3mL無(wu)20×洗(xi)滌(di)(di)緩(huan)沖(chong)液20mL20mL按(an)(an)說(shuo)明(ming)書(shu)(shu)進行(xing)稀釋底(di)物(wu)(wu)(wu) A6mL3mL無(wu)底(di)物(wu)(wu)(wu) B6mL3mL無(wu)終止液6mL3mL無(wu)封(feng)板(ban)(ban)(ban)膜2 張2 張無(wu)說(shuo)明(ming)書(shu)(shu)1 份1 份無(wu)自封(feng)袋1 個(ge)(ge)1 個(ge)(ge)無(wu) 操(cao)作(zuo)(zuo)步驟1. 準備(bei)(bei)工作(zuo)(zuo):將(jiang)試(shi)劑盒(he)從(cong)冰箱中(zhong)取(qu)出(chu)(chu),室溫(wen)(wen)(wen)放置(zhi)(zhi)30分(fen)鐘(zhong),使試(shi)劑盒(he)恢(hui)復(fu)至室溫(wen)(wen)(wen)。同時(shi)(shi)(shi)準備(bei)(bei)好(hao)實(shi)(shi)驗(yan)所(suo)需(xu)(xu)的(de)(de)(de)材料和(he)(he)器具。2. 加(jia)(jia)(jia)(jia)(jia)樣(yang)(yang):在(zai)酶(mei)(mei)(mei)(mei)標(biao)板(ban)(ban)(ban)的(de)(de)(de)弟1個(ge)(ge)孔(kong)(kong)中(zhong)加(jia)(jia)(jia)(jia)(jia)入(ru)標(biao)準品(pin)(pin)50μL,然后在(zai)后續的(de)(de)(de)孔(kong)(kong)中(zhong)依次加(jia)(jia)(jia)(jia)(jia)入(ru)待(dai)測樣(yang)(yang)本50μL。3. 溫(wen)(wen)(wen)育:將(jiang)酶(mei)(mei)(mei)(mei)標(biao)板(ban)(ban)(ban)密(mi)封(feng),在(zai)37℃下(xia)溫(wen)(wen)(wen)育30分(fen)鐘(zhong)。4. 洗(xi)滌(di)(di):用(yong)(yong)洗(xi)滌(di)(di)液清洗(xi)酶(mei)(mei)(mei)(mei)標(biao)板(ban)(ban)(ban),共洗(xi)滌(di)(di)5次。每(mei)次洗(xi)滌(di)(di)后,用(yong)(yong)吸水(shui)(shui)紙輕(qing)(qing)輕(qing)(qing)吸干(gan)(gan)孔(kong)(kong)內液體(ti)。5. 加(jia)(jia)(jia)(jia)(jia)酶(mei)(mei)(mei)(mei):在(zai)每(mei)個(ge)(ge)孔(kong)(kong)中(zhong)加(jia)(jia)(jia)(jia)(jia)入(ru)酶(mei)(mei)(mei)(mei)標(biao)物(wu)(wu)(wu)100μL。6. 溫(wen)(wen)(wen)育:再次將(jiang)酶(mei)(mei)(mei)(mei)標(biao)板(ban)(ban)(ban)密(mi)封(feng),在(zai)37℃下(xia)溫(wen)(wen)(wen)育30分(fen)鐘(zhong)。7. 洗(xi)滌(di)(di):用(yong)(yong)洗(xi)滌(di)(di)液清洗(xi)酶(mei)(mei)(mei)(mei)標(biao)板(ban)(ban)(ban),共洗(xi)滌(di)(di)5次。每(mei)次洗(xi)滌(di)(di)后,用(yong)(yong)吸水(shui)(shui)紙輕(qing)(qing)輕(qing)(qing)吸干(gan)(gan)孔(kong)(kong)內液體(ti)。8. 加(jia)(jia)(jia)(jia)(jia)底(di)物(wu)(wu)(wu):在(zai)每(mei)個(ge)(ge)孔(kong)(kong)中(zhong)加(jia)(jia)(jia)(jia)(jia)入(ru)底(di)物(wu)(wu)(wu)A液50μL,再加(jia)(jia)(jia)(jia)(jia)入(ru)底(di)物(wu)(wu)(wu)B液50μL。9. 終止反應:在(zai)每(mei)個(ge)(ge)孔(kong)(kong)中(zhong)加(jia)(jia)(jia)(jia)(jia)入(ru)終止液50μL。10. 檢(jian)測:用(yong)(yong)酶(mei)(mei)(mei)(mei)標(biao)儀在(zai)450nm波(bo)長下(xia)讀取(qu)各孔(kong)(kong)的(de)(de)(de)吸光(guang)度(du)(du)(du)(OD值)。 結果分(fen)析根據標(biao)準品(pin)(pin)和(he)(he)樣(yang)(yang)本的(de)(de)(de)OD值,繪制(zhi)(zhi)標(biao)準曲線,從(cong)而計算出(chu)(chu)樣(yang)(yang)本中(zhong)角蛋(dan)白(bai)1(KRT1)的(de)(de)(de)濃度(du)(du)(du)。具體(ti)分(fen)析方法可參考試(shi)劑盒(he)說(shuo)明(ming)書(shu)(shu)或相關文獻。注意事項(xiang)1. 在(zai)操(cao)作(zuo)(zuo)過程中(zhong),應保(bao)持(chi)酶(mei)(mei)(mei)(mei)標(biao)板(ban)(ban)(ban)的(de)(de)(de)干(gan)(gan)燥,避(bi)免水(shui)(shui)分(fen)進入(ru)孔(kong)(kong)內。2. 加(jia)(jia)(jia)(jia)(jia)樣(yang)(yang)時(shi)(shi)(shi)需(xu)(xu)確(que)(que)保(bao)加(jia)(jia)(jia)(jia)(jia)樣(yang)(yang)準確(que)(que)、無(wu)氣泡(pao)產生。3. 溫(wen)(wen)(wen)育和(he)(he)洗(xi)滌(di)(di)過程中(zhong)需(xu)(xu)嚴格控(kong)制(zhi)(zhi)時(shi)(shi)(shi)間和(he)(he)溫(wen)(wen)(wen)度(du)(du)(du),確(que)(que)保(bao)實(shi)(shi)驗(yan)結果的(de)(de)(de)準確(que)(que)性。4. 在(zai)使用(yong)(yong)酶(mei)(mei)(mei)(mei)標(biao)儀讀取(qu)OD值時(shi)(shi)(shi),需(xu)(xu)確(que)(que)保(bao)波(bo)長設置(zhi)(zhi)正確(que)(que),避(bi)免誤差產生。5. 在(zai)整個(ge)(ge)實(shi)(shi)驗(yan)過程中(zhong),需(xu)(xu)保(bao)持(chi)操(cao)作(zuo)(zuo)環境的(de)(de)(de)清潔和(he)(he)整潔,避(bi)免交(jiao)叉(cha)污染。
猴核糖體(ti)(ti)蛋(dan)(dan)白S6激酶(mei)(mei)(mei)β1(RPS6Kβ1)酶(mei)(mei)(mei)聯免(mian)(mian)疫試(shi)劑盒(he)實(shi)驗原理(li) ELISA運用(yong)了(le)免(mian)(mian)疫學原理(li)和(he)(he)化學反應顯色(se)原理(li),需(xu)(xu)(xu)要將抗(kang)(kang)原或抗(kang)(kang)體(ti)(ti)預(yu)包被在(zai)(zai)(zai)(zai)(zai)固相載(zai)體(ti)(ti)表面,使(shi)后來形成的(de)(de)(de)抗(kang)(kang)原-抗(kang)(kang)體(ti)(ti)-酶(mei)(mei)(mei) (熒光(guang)基(ji)團) -底(di)物(wu)復(fu)合物(wu)黏(nian)附在(zai)(zai)(zai)(zai)(zai)載(zai)體(ti)(ti)上,通過檢(jian)測(ce)OD值或發光(guang)值,來檢(jian)測(ce)靶蛋(dan)(dan)白的(de)(de)(de)含量(liang)。 實(shi)驗所需(xu)(xu)(xu)自備(bei)(bei)物(wu)品1. 酶(mei)(mei)(mei)標(biao)(biao)(biao)(biao)儀(yi)(450nm)2. 加(jia)(jia)樣(yang)(yang)器及(ji)槍頭:0.5-10uL、2-20uL、20-200uL、200-1000uL3. 37℃恒溫(wen)(wen)箱 試(shi)劑盒(he)組成名稱96 孔(kong)配置48 孔(kong)配置備(bei)(bei)注微孔(kong)酶(mei)(mei)(mei)標(biao)(biao)(biao)(biao)板(ban)12 孔(kong)×8 條(tiao)12 孔(kong)×4 條(tiao)無(wu)(wu)標(biao)(biao)(biao)(biao)準(zhun)品0.5mL0.5mL按(an)(an)說(shuo)(shuo)明(ming)(ming)書(shu)(shu)進行(xing)稀(xi)釋(shi)標(biao)(biao)(biao)(biao)準(zhun)品稀(xi)釋(shi)液(ye)(ye)(ye)(ye)6mL3mL按(an)(an)說(shuo)(shuo)明(ming)(ming)書(shu)(shu)進行(xing)稀(xi)釋(shi)樣(yang)(yang)本(ben)稀(xi)釋(shi)液(ye)(ye)(ye)(ye)6mL3mL按(an)(an)說(shuo)(shuo)明(ming)(ming)書(shu)(shu)進行(xing)稀(xi)釋(shi)檢(jian)測(ce)抗(kang)(kang)體(ti)(ti)-HRP6mL3mL無(wu)(wu)20×洗(xi)(xi)(xi)(xi)滌(di)(di)(di)(di)緩沖液(ye)(ye)(ye)(ye)20mL20mL按(an)(an)說(shuo)(shuo)明(ming)(ming)書(shu)(shu)進行(xing)稀(xi)釋(shi)底(di)物(wu) A6mL3mL無(wu)(wu)底(di)物(wu) B6mL3mL無(wu)(wu)終止(zhi)液(ye)(ye)(ye)(ye)6mL3mL無(wu)(wu)封(feng)板(ban)膜2 張2 張無(wu)(wu)說(shuo)(shuo)明(ming)(ming)書(shu)(shu)1 份1 份無(wu)(wu)自封(feng)袋1 個(ge)(ge)1 個(ge)(ge)無(wu)(wu) 操作步驟(zou)1. 準(zhun)備(bei)(bei)工(gong)作:將試(shi)劑盒(he)從(cong)冰箱中(zhong)(zhong)(zhong)取(qu)出(chu),室溫(wen)(wen)放置30分鐘,使(shi)試(shi)劑盒(he)恢復(fu)至(zhi)室溫(wen)(wen)。同時(shi)(shi)準(zhun)備(bei)(bei)好實(shi)驗所需(xu)(xu)(xu)的(de)(de)(de)材料和(he)(he)器具。2. 加(jia)(jia)樣(yang)(yang):在(zai)(zai)(zai)(zai)(zai)酶(mei)(mei)(mei)標(biao)(biao)(biao)(biao)板(ban)的(de)(de)(de)弟1個(ge)(ge)孔(kong)中(zhong)(zhong)(zhong)加(jia)(jia)入(ru)標(biao)(biao)(biao)(biao)準(zhun)品50μL,然后在(zai)(zai)(zai)(zai)(zai)后續(xu)的(de)(de)(de)孔(kong)中(zhong)(zhong)(zhong)依次(ci)加(jia)(jia)入(ru)待測(ce)樣(yang)(yang)本(ben)50μL。3. 溫(wen)(wen)育:將酶(mei)(mei)(mei)標(biao)(biao)(biao)(biao)板(ban)密封(feng),在(zai)(zai)(zai)(zai)(zai)37℃下溫(wen)(wen)育30分鐘。4. 洗(xi)(xi)(xi)(xi)滌(di)(di)(di)(di):用(yong)洗(xi)(xi)(xi)(xi)滌(di)(di)(di)(di)液(ye)(ye)(ye)(ye)清洗(xi)(xi)(xi)(xi)酶(mei)(mei)(mei)標(biao)(biao)(biao)(biao)板(ban),共洗(xi)(xi)(xi)(xi)滌(di)(di)(di)(di)5次(ci)。每(mei)(mei)(mei)次(ci)洗(xi)(xi)(xi)(xi)滌(di)(di)(di)(di)后,用(yong)吸水(shui)(shui)紙(zhi)輕(qing)輕(qing)吸干孔(kong)內(nei)液(ye)(ye)(ye)(ye)體(ti)(ti)。5. 加(jia)(jia)酶(mei)(mei)(mei):在(zai)(zai)(zai)(zai)(zai)每(mei)(mei)(mei)個(ge)(ge)孔(kong)中(zhong)(zhong)(zhong)加(jia)(jia)入(ru)酶(mei)(mei)(mei)標(biao)(biao)(biao)(biao)物(wu)100μL。6. 溫(wen)(wen)育:再次(ci)將酶(mei)(mei)(mei)標(biao)(biao)(biao)(biao)板(ban)密封(feng),在(zai)(zai)(zai)(zai)(zai)37℃下溫(wen)(wen)育30分鐘。7. 洗(xi)(xi)(xi)(xi)滌(di)(di)(di)(di):用(yong)洗(xi)(xi)(xi)(xi)滌(di)(di)(di)(di)液(ye)(ye)(ye)(ye)清洗(xi)(xi)(xi)(xi)酶(mei)(mei)(mei)標(biao)(biao)(biao)(biao)板(ban),共洗(xi)(xi)(xi)(xi)滌(di)(di)(di)(di)5次(ci)。每(mei)(mei)(mei)次(ci)洗(xi)(xi)(xi)(xi)滌(di)(di)(di)(di)后,用(yong)吸水(shui)(shui)紙(zhi)輕(qing)輕(qing)吸干孔(kong)內(nei)液(ye)(ye)(ye)(ye)體(ti)(ti)。8. 加(jia)(jia)底(di)物(wu):在(zai)(zai)(zai)(zai)(zai)每(mei)(mei)(mei)個(ge)(ge)孔(kong)中(zhong)(zhong)(zhong)加(jia)(jia)入(ru)底(di)物(wu)A液(ye)(ye)(ye)(ye)50μL,再加(jia)(jia)入(ru)底(di)物(wu)B液(ye)(ye)(ye)(ye)50μL。9. 終止(zhi)反應:在(zai)(zai)(zai)(zai)(zai)每(mei)(mei)(mei)個(ge)(ge)孔(kong)中(zhong)(zhong)(zhong)加(jia)(jia)入(ru)終止(zhi)液(ye)(ye)(ye)(ye)50μL。10. 檢(jian)測(ce):用(yong)酶(mei)(mei)(mei)標(biao)(biao)(biao)(biao)儀(yi)在(zai)(zai)(zai)(zai)(zai)450nm波長下讀取(qu)各孔(kong)的(de)(de)(de)吸光(guang)度(du)(du)(OD值)。 結果分析根據標(biao)(biao)(biao)(biao)準(zhun)品和(he)(he)樣(yang)(yang)本(ben)的(de)(de)(de)OD值,繪制(zhi)(zhi)標(biao)(biao)(biao)(biao)準(zhun)曲(qu)線,從(cong)而計算出(chu)樣(yang)(yang)本(ben)中(zhong)(zhong)(zhong)角蛋(dan)(dan)白1(KRT1)的(de)(de)(de)濃度(du)(du)。具體(ti)(ti)分析方(fang)法(fa)可參(can)考試(shi)劑盒(he)說(shuo)(shuo)明(ming)(ming)書(shu)(shu)或相關文獻(xian)。注意事項1. 在(zai)(zai)(zai)(zai)(zai)操作過程(cheng)中(zhong)(zhong)(zhong),應保(bao)持酶(mei)(mei)(mei)標(biao)(biao)(biao)(biao)板(ban)的(de)(de)(de)干燥,避免(mian)(mian)水(shui)(shui)分進入(ru)孔(kong)內(nei)。2. 加(jia)(jia)樣(yang)(yang)時(shi)(shi)需(xu)(xu)(xu)確(que)保(bao)加(jia)(jia)樣(yang)(yang)準(zhun)確(que)、無(wu)(wu)氣泡產生。3. 溫(wen)(wen)育和(he)(he)洗(xi)(xi)(xi)(xi)滌(di)(di)(di)(di)過程(cheng)中(zhong)(zhong)(zhong)需(xu)(xu)(xu)嚴格控制(zhi)(zhi)時(shi)(shi)間(jian)和(he)(he)溫(wen)(wen)度(du)(du),確(que)保(bao)實(shi)驗結果的(de)(de)(de)準(zhun)確(que)性。4. 在(zai)(zai)(zai)(zai)(zai)使(shi)用(yong)酶(mei)(mei)(mei)標(biao)(biao)(biao)(biao)儀(yi)讀取(qu)OD值時(shi)(shi),需(xu)(xu)(xu)確(que)保(bao)波長設置正(zheng)確(que),避免(mian)(mian)誤(wu)差產生。5. 在(zai)(zai)(zai)(zai)(zai)整(zheng)個(ge)(ge)實(shi)驗過程(cheng)中(zhong)(zhong)(zhong),需(xu)(xu)(xu)保(bao)持操作環境的(de)(de)(de)清潔(jie)和(he)(he)整(zheng)潔(jie),避免(mian)(mian)交(jiao)叉污(wu)染。
猴血管(guan)緊(jin)張素轉化(hua)酶(mei)2(ACE2)酶(mei)聯(lian)免(mian)疫試(shi)(shi)劑(ji)盒(he) 實(shi)(shi)驗(yan)原理(li) ELISA運用(yong)了免(mian)疫學(xue)原理(li)和(he)化(hua)學(xue)反應顯色原理(li),需(xu)要將(jiang)抗(kang)(kang)原或抗(kang)(kang)體(ti)預包(bao)被在(zai)(zai)固(gu)相載體(ti)表面,使后來形成(cheng)的(de)(de)(de)(de)抗(kang)(kang)原-抗(kang)(kang)體(ti)-酶(mei) (熒(ying)光基團(tuan)) -底(di)物(wu)(wu)(wu)(wu)復合(he)物(wu)(wu)(wu)(wu)黏附在(zai)(zai)載體(ti)上,通過(guo)檢(jian)(jian)測OD值(zhi)或發光值(zhi),來檢(jian)(jian)測靶蛋(dan)白的(de)(de)(de)(de)含量。 實(shi)(shi)驗(yan)所需(xu)自備(bei)(bei)物(wu)(wu)(wu)(wu)品1. 酶(mei)標(biao)儀(yi)(450nm)2. 加(jia)樣(yang)(yang)(yang)器及槍頭:0.5-10uL、2-20uL、20-200uL、200-1000uL3. 37℃恒溫(wen)箱(xiang) 試(shi)(shi)劑(ji)盒(he)組成(cheng)名稱96 孔(kong)(kong)(kong)(kong)配置48 孔(kong)(kong)(kong)(kong)配置備(bei)(bei)注(zhu)微孔(kong)(kong)(kong)(kong)酶(mei)標(biao)板(ban)(ban)(ban)(ban)12 孔(kong)(kong)(kong)(kong)×8 條(tiao)12 孔(kong)(kong)(kong)(kong)×4 條(tiao)無標(biao)準品0.5mL0.5mL按說(shuo)明書(shu)進行稀(xi)釋(shi)標(biao)準品稀(xi)釋(shi)液(ye)(ye)(ye)6mL3mL按說(shuo)明書(shu)進行稀(xi)釋(shi)樣(yang)(yang)(yang)本稀(xi)釋(shi)液(ye)(ye)(ye)6mL3mL按說(shuo)明書(shu)進行稀(xi)釋(shi)檢(jian)(jian)測抗(kang)(kang)體(ti)-HRP6mL3mL無20×洗(xi)滌(di)(di)(di)(di)(di)緩沖液(ye)(ye)(ye)20mL20mL按說(shuo)明書(shu)進行稀(xi)釋(shi)底(di)物(wu)(wu)(wu)(wu) A6mL3mL無底(di)物(wu)(wu)(wu)(wu) B6mL3mL無終止液(ye)(ye)(ye)6mL3mL無封板(ban)(ban)(ban)(ban)膜2 張2 張無說(shuo)明書(shu)1 份(fen)1 份(fen)無自封袋1 個(ge)1 個(ge)無 操作步驟(zou)1. 準備(bei)(bei)工(gong)作:將(jiang)試(shi)(shi)劑(ji)盒(he)從冰箱(xiang)中(zhong)取(qu)出,室溫(wen)放(fang)置30分(fen)(fen)鐘(zhong),使試(shi)(shi)劑(ji)盒(he)恢復至室溫(wen)。同時(shi)準備(bei)(bei)好(hao)實(shi)(shi)驗(yan)所需(xu)的(de)(de)(de)(de)材料和(he)器具。2. 加(jia)樣(yang)(yang)(yang):在(zai)(zai)酶(mei)標(biao)板(ban)(ban)(ban)(ban)的(de)(de)(de)(de)弟1個(ge)孔(kong)(kong)(kong)(kong)中(zhong)加(jia)入(ru)(ru)標(biao)準品50μL,然后在(zai)(zai)后續的(de)(de)(de)(de)孔(kong)(kong)(kong)(kong)中(zhong)依次加(jia)入(ru)(ru)待測樣(yang)(yang)(yang)本50μL。3. 溫(wen)育(yu)(yu):將(jiang)酶(mei)標(biao)板(ban)(ban)(ban)(ban)密封,在(zai)(zai)37℃下(xia)溫(wen)育(yu)(yu)30分(fen)(fen)鐘(zhong)。4. 洗(xi)滌(di)(di)(di)(di)(di):用(yong)洗(xi)滌(di)(di)(di)(di)(di)液(ye)(ye)(ye)清洗(xi)酶(mei)標(biao)板(ban)(ban)(ban)(ban),共(gong)洗(xi)滌(di)(di)(di)(di)(di)5次。每次洗(xi)滌(di)(di)(di)(di)(di)后,用(yong)吸水(shui)紙輕輕吸干(gan)孔(kong)(kong)(kong)(kong)內液(ye)(ye)(ye)體(ti)。5. 加(jia)酶(mei):在(zai)(zai)每個(ge)孔(kong)(kong)(kong)(kong)中(zhong)加(jia)入(ru)(ru)酶(mei)標(biao)物(wu)(wu)(wu)(wu)100μL。6. 溫(wen)育(yu)(yu):再次將(jiang)酶(mei)標(biao)板(ban)(ban)(ban)(ban)密封,在(zai)(zai)37℃下(xia)溫(wen)育(yu)(yu)30分(fen)(fen)鐘(zhong)。7. 洗(xi)滌(di)(di)(di)(di)(di):用(yong)洗(xi)滌(di)(di)(di)(di)(di)液(ye)(ye)(ye)清洗(xi)酶(mei)標(biao)板(ban)(ban)(ban)(ban),共(gong)洗(xi)滌(di)(di)(di)(di)(di)5次。每次洗(xi)滌(di)(di)(di)(di)(di)后,用(yong)吸水(shui)紙輕輕吸干(gan)孔(kong)(kong)(kong)(kong)內液(ye)(ye)(ye)體(ti)。8. 加(jia)底(di)物(wu)(wu)(wu)(wu):在(zai)(zai)每個(ge)孔(kong)(kong)(kong)(kong)中(zhong)加(jia)入(ru)(ru)底(di)物(wu)(wu)(wu)(wu)A液(ye)(ye)(ye)50μL,再加(jia)入(ru)(ru)底(di)物(wu)(wu)(wu)(wu)B液(ye)(ye)(ye)50μL。9. 終止反應:在(zai)(zai)每個(ge)孔(kong)(kong)(kong)(kong)中(zhong)加(jia)入(ru)(ru)終止液(ye)(ye)(ye)50μL。10. 檢(jian)(jian)測:用(yong)酶(mei)標(biao)儀(yi)在(zai)(zai)450nm波(bo)長下(xia)讀(du)(du)取(qu)各孔(kong)(kong)(kong)(kong)的(de)(de)(de)(de)吸光度(OD值(zhi))。 結果分(fen)(fen)析根據(ju)標(biao)準品和(he)樣(yang)(yang)(yang)本的(de)(de)(de)(de)OD值(zhi),繪制標(biao)準曲線(xian),從而計算出樣(yang)(yang)(yang)本中(zhong)角蛋(dan)白1(KRT1)的(de)(de)(de)(de)濃度。具體(ti)分(fen)(fen)析方法(fa)可(ke)參考試(shi)(shi)劑(ji)盒(he)說(shuo)明書(shu)或相關文獻。注(zhu)意事項1. 在(zai)(zai)操作過(guo)程(cheng)中(zhong),應保(bao)持酶(mei)標(biao)板(ban)(ban)(ban)(ban)的(de)(de)(de)(de)干(gan)燥,避(bi)(bi)免(mian)水(shui)分(fen)(fen)進入(ru)(ru)孔(kong)(kong)(kong)(kong)內。2. 加(jia)樣(yang)(yang)(yang)時(shi)需(xu)確(que)保(bao)加(jia)樣(yang)(yang)(yang)準確(que)、無氣泡(pao)產生。3. 溫(wen)育(yu)(yu)和(he)洗(xi)滌(di)(di)(di)(di)(di)過(guo)程(cheng)中(zhong)需(xu)嚴格控制時(shi)間和(he)溫(wen)度,確(que)保(bao)實(shi)(shi)驗(yan)結果的(de)(de)(de)(de)準確(que)性。4. 在(zai)(zai)使用(yong)酶(mei)標(biao)儀(yi)讀(du)(du)取(qu)OD值(zhi)時(shi),需(xu)確(que)保(bao)波(bo)長設(she)置正確(que),避(bi)(bi)免(mian)誤差產生。5. 在(zai)(zai)整個(ge)實(shi)(shi)驗(yan)過(guo)程(cheng)中(zhong),需(xu)保(bao)持操作環境的(de)(de)(de)(de)清潔和(he)整潔,避(bi)(bi)免(mian)交叉污染。
猴細胞周期素D1(CCND1)酶(mei)(mei)(mei)聯免疫試(shi)(shi)劑(ji)盒(he) 實驗(yan)(yan)原(yuan)(yuan)理 ELISA運用(yong)了免疫學原(yuan)(yuan)理和(he)化學反應顯(xian)色原(yuan)(yuan)理,需要將抗(kang)原(yuan)(yuan)或抗(kang)體預包被在(zai)(zai)(zai)固相載體表面(mian),使后(hou)(hou)來(lai)形成的(de)(de)(de)抗(kang)原(yuan)(yuan)-抗(kang)體-酶(mei)(mei)(mei) (熒光(guang)基團) -底物(wu)(wu)(wu)復合物(wu)(wu)(wu)黏附在(zai)(zai)(zai)載體上(shang),通過(guo)檢(jian)測(ce)OD值(zhi)或發光(guang)值(zhi),來(lai)檢(jian)測(ce)靶蛋白的(de)(de)(de)含(han)量。 實驗(yan)(yan)所(suo)需自備(bei)物(wu)(wu)(wu)品(pin)1. 酶(mei)(mei)(mei)標(biao)儀(450nm)2. 加(jia)(jia)(jia)樣器及槍頭(tou):0.5-10uL、2-20uL、20-200uL、200-1000uL3. 37℃恒(heng)溫(wen)(wen)(wen)箱 試(shi)(shi)劑(ji)盒(he)組(zu)成名稱96 孔(kong)(kong)配(pei)置48 孔(kong)(kong)配(pei)置備(bei)注微(wei)孔(kong)(kong)酶(mei)(mei)(mei)標(biao)板(ban)12 孔(kong)(kong)×8 條12 孔(kong)(kong)×4 條無(wu)標(biao)準(zhun)品(pin)0.5mL0.5mL按(an)(an)(an)說(shuo)明書(shu)進(jin)行(xing)(xing)稀釋(shi)標(biao)準(zhun)品(pin)稀釋(shi)液(ye)(ye)(ye)(ye)6mL3mL按(an)(an)(an)說(shuo)明書(shu)進(jin)行(xing)(xing)稀釋(shi)樣本稀釋(shi)液(ye)(ye)(ye)(ye)6mL3mL按(an)(an)(an)說(shuo)明書(shu)進(jin)行(xing)(xing)稀釋(shi)檢(jian)測(ce)抗(kang)體-HRP6mL3mL無(wu)20×洗(xi)(xi)(xi)滌(di)緩沖液(ye)(ye)(ye)(ye)20mL20mL按(an)(an)(an)說(shuo)明書(shu)進(jin)行(xing)(xing)稀釋(shi)底物(wu)(wu)(wu) A6mL3mL無(wu)底物(wu)(wu)(wu) B6mL3mL無(wu)終止液(ye)(ye)(ye)(ye)6mL3mL無(wu)封板(ban)膜2 張2 張無(wu)說(shuo)明書(shu)1 份1 份無(wu)自封袋1 個(ge)1 個(ge)無(wu) 操作(zuo)(zuo)步驟(zou)1. 準(zhun)備(bei)工(gong)作(zuo)(zuo):將試(shi)(shi)劑(ji)盒(he)從冰(bing)箱中(zhong)(zhong)取(qu)出(chu),室溫(wen)(wen)(wen)放置30分(fen)鐘(zhong)(zhong),使試(shi)(shi)劑(ji)盒(he)恢復至(zhi)室溫(wen)(wen)(wen)。同時準(zhun)備(bei)好實驗(yan)(yan)所(suo)需的(de)(de)(de)材料和(he)器具。2. 加(jia)(jia)(jia)樣:在(zai)(zai)(zai)酶(mei)(mei)(mei)標(biao)板(ban)的(de)(de)(de)弟1個(ge)孔(kong)(kong)中(zhong)(zhong)加(jia)(jia)(jia)入(ru)標(biao)準(zhun)品(pin)50μL,然后(hou)(hou)在(zai)(zai)(zai)后(hou)(hou)續的(de)(de)(de)孔(kong)(kong)中(zhong)(zhong)依次(ci)加(jia)(jia)(jia)入(ru)待測(ce)樣本50μL。3. 溫(wen)(wen)(wen)育(yu):將酶(mei)(mei)(mei)標(biao)板(ban)密(mi)封,在(zai)(zai)(zai)37℃下溫(wen)(wen)(wen)育(yu)30分(fen)鐘(zhong)(zhong)。4. 洗(xi)(xi)(xi)滌(di):用(yong)洗(xi)(xi)(xi)滌(di)液(ye)(ye)(ye)(ye)清洗(xi)(xi)(xi)酶(mei)(mei)(mei)標(biao)板(ban),共洗(xi)(xi)(xi)滌(di)5次(ci)。每(mei)次(ci)洗(xi)(xi)(xi)滌(di)后(hou)(hou),用(yong)吸(xi)(xi)水紙輕(qing)(qing)(qing)輕(qing)(qing)(qing)吸(xi)(xi)干孔(kong)(kong)內(nei)液(ye)(ye)(ye)(ye)體。5. 加(jia)(jia)(jia)酶(mei)(mei)(mei):在(zai)(zai)(zai)每(mei)個(ge)孔(kong)(kong)中(zhong)(zhong)加(jia)(jia)(jia)入(ru)酶(mei)(mei)(mei)標(biao)物(wu)(wu)(wu)100μL。6. 溫(wen)(wen)(wen)育(yu):再次(ci)將酶(mei)(mei)(mei)標(biao)板(ban)密(mi)封,在(zai)(zai)(zai)37℃下溫(wen)(wen)(wen)育(yu)30分(fen)鐘(zhong)(zhong)。7. 洗(xi)(xi)(xi)滌(di):用(yong)洗(xi)(xi)(xi)滌(di)液(ye)(ye)(ye)(ye)清洗(xi)(xi)(xi)酶(mei)(mei)(mei)標(biao)板(ban),共洗(xi)(xi)(xi)滌(di)5次(ci)。每(mei)次(ci)洗(xi)(xi)(xi)滌(di)后(hou)(hou),用(yong)吸(xi)(xi)水紙輕(qing)(qing)(qing)輕(qing)(qing)(qing)吸(xi)(xi)干孔(kong)(kong)內(nei)液(ye)(ye)(ye)(ye)體。8. 加(jia)(jia)(jia)底物(wu)(wu)(wu):在(zai)(zai)(zai)每(mei)個(ge)孔(kong)(kong)中(zhong)(zhong)加(jia)(jia)(jia)入(ru)底物(wu)(wu)(wu)A液(ye)(ye)(ye)(ye)50μL,再加(jia)(jia)(jia)入(ru)底物(wu)(wu)(wu)B液(ye)(ye)(ye)(ye)50μL。9. 終止反應:在(zai)(zai)(zai)每(mei)個(ge)孔(kong)(kong)中(zhong)(zhong)加(jia)(jia)(jia)入(ru)終止液(ye)(ye)(ye)(ye)50μL。10. 檢(jian)測(ce):用(yong)酶(mei)(mei)(mei)標(biao)儀在(zai)(zai)(zai)450nm波長下讀(du)取(qu)各(ge)孔(kong)(kong)的(de)(de)(de)吸(xi)(xi)光(guang)度(du)(OD值(zhi))。 結果(guo)分(fen)析根(gen)據標(biao)準(zhun)品(pin)和(he)樣本的(de)(de)(de)OD值(zhi),繪制標(biao)準(zhun)曲線,從而計算出(chu)樣本中(zhong)(zhong)角(jiao)蛋白1(KRT1)的(de)(de)(de)濃度(du)。具體分(fen)析方法可(ke)參考試(shi)(shi)劑(ji)盒(he)說(shuo)明書(shu)或相關(guan)文(wen)獻。注意事項(xiang)1. 在(zai)(zai)(zai)操作(zuo)(zuo)過(guo)程(cheng)中(zhong)(zhong),應保持(chi)(chi)酶(mei)(mei)(mei)標(biao)板(ban)的(de)(de)(de)干燥,避免水分(fen)進(jin)入(ru)孔(kong)(kong)內(nei)。2. 加(jia)(jia)(jia)樣時需確(que)保加(jia)(jia)(jia)樣準(zhun)確(que)、無(wu)氣泡(pao)產(chan)(chan)生(sheng)。3. 溫(wen)(wen)(wen)育(yu)和(he)洗(xi)(xi)(xi)滌(di)過(guo)程(cheng)中(zhong)(zhong)需嚴格控(kong)制時間和(he)溫(wen)(wen)(wen)度(du),確(que)保實驗(yan)(yan)結果(guo)的(de)(de)(de)準(zhun)確(que)性。4. 在(zai)(zai)(zai)使用(yong)酶(mei)(mei)(mei)標(biao)儀讀(du)取(qu)OD值(zhi)時,需確(que)保波長設置正確(que),避免誤(wu)差產(chan)(chan)生(sheng)。5. 在(zai)(zai)(zai)整個(ge)實驗(yan)(yan)過(guo)程(cheng)中(zhong)(zhong),需保持(chi)(chi)操作(zuo)(zuo)環(huan)境的(de)(de)(de)清潔和(he)整潔,避免交(jiao)叉污(wu)染。
猴重組(zu)激活基(ji)因1(RAG-1)酶(mei)(mei)(mei)(mei)聯免(mian)疫試(shi)劑(ji)盒 實驗原(yuan)理(li) ELISA運用(yong)了免(mian)疫學(xue)原(yuan)理(li)和(he)化(hua)學(xue)反應顯色原(yuan)理(li),需(xu)(xu)(xu)要將(jiang)抗原(yuan)或抗體(ti)(ti)預包被(bei)在(zai)(zai)(zai)固(gu)相(xiang)載體(ti)(ti)表面(mian),使(shi)后(hou)來形成的(de)(de)(de)抗原(yuan)-抗體(ti)(ti)-酶(mei)(mei)(mei)(mei) (熒光(guang)基(ji)團(tuan)) -底(di)物(wu)(wu)(wu)(wu)復合物(wu)(wu)(wu)(wu)黏(nian)附在(zai)(zai)(zai)載體(ti)(ti)上,通過檢測(ce)OD值或發光(guang)值,來檢測(ce)靶(ba)蛋白(bai)的(de)(de)(de)含量。 實驗所需(xu)(xu)(xu)自備(bei)(bei)物(wu)(wu)(wu)(wu)品1. 酶(mei)(mei)(mei)(mei)標(biao)儀(450nm)2. 加樣(yang)器及槍頭:0.5-10uL、2-20uL、20-200uL、200-1000uL3. 37℃恒溫(wen)箱 試(shi)劑(ji)盒組(zu)成名稱96 孔(kong)(kong)(kong)(kong)(kong)配置(zhi)48 孔(kong)(kong)(kong)(kong)(kong)配置(zhi)備(bei)(bei)注微孔(kong)(kong)(kong)(kong)(kong)酶(mei)(mei)(mei)(mei)標(biao)板(ban)12 孔(kong)(kong)(kong)(kong)(kong)×8 條12 孔(kong)(kong)(kong)(kong)(kong)×4 條無(wu)標(biao)準(zhun)(zhun)品0.5mL0.5mL按說明書(shu)(shu)進(jin)行(xing)稀(xi)(xi)釋(shi)標(biao)準(zhun)(zhun)品稀(xi)(xi)釋(shi)液(ye)(ye)6mL3mL按說明書(shu)(shu)進(jin)行(xing)稀(xi)(xi)釋(shi)樣(yang)本稀(xi)(xi)釋(shi)液(ye)(ye)6mL3mL按說明書(shu)(shu)進(jin)行(xing)稀(xi)(xi)釋(shi)檢測(ce)抗體(ti)(ti)-HRP6mL3mL無(wu)20×洗滌(di)(di)緩沖液(ye)(ye)20mL20mL按說明書(shu)(shu)進(jin)行(xing)稀(xi)(xi)釋(shi)底(di)物(wu)(wu)(wu)(wu) A6mL3mL無(wu)底(di)物(wu)(wu)(wu)(wu) B6mL3mL無(wu)終止液(ye)(ye)6mL3mL無(wu)封(feng)板(ban)膜2 張2 張無(wu)說明書(shu)(shu)1 份(fen)(fen)1 份(fen)(fen)無(wu)自封(feng)袋(dai)1 個(ge)1 個(ge)無(wu) 操作步驟1. 準(zhun)(zhun)備(bei)(bei)工作:將(jiang)試(shi)劑(ji)盒從冰箱中(zhong)(zhong)取(qu)出(chu),室溫(wen)放置(zhi)30分(fen)(fen)鐘,使(shi)試(shi)劑(ji)盒恢(hui)復至室溫(wen)。同時(shi)準(zhun)(zhun)備(bei)(bei)好實驗所需(xu)(xu)(xu)的(de)(de)(de)材料(liao)和(he)器具。2. 加樣(yang):在(zai)(zai)(zai)酶(mei)(mei)(mei)(mei)標(biao)板(ban)的(de)(de)(de)弟1個(ge)孔(kong)(kong)(kong)(kong)(kong)中(zhong)(zhong)加入標(biao)準(zhun)(zhun)品50μL,然(ran)后(hou)在(zai)(zai)(zai)后(hou)續的(de)(de)(de)孔(kong)(kong)(kong)(kong)(kong)中(zhong)(zhong)依(yi)次加入待測(ce)樣(yang)本50μL。3. 溫(wen)育(yu):將(jiang)酶(mei)(mei)(mei)(mei)標(biao)板(ban)密封(feng),在(zai)(zai)(zai)37℃下(xia)溫(wen)育(yu)30分(fen)(fen)鐘。4. 洗滌(di)(di):用(yong)洗滌(di)(di)液(ye)(ye)清(qing)(qing)洗酶(mei)(mei)(mei)(mei)標(biao)板(ban),共洗滌(di)(di)5次。每(mei)次洗滌(di)(di)后(hou),用(yong)吸(xi)水(shui)紙輕輕吸(xi)干孔(kong)(kong)(kong)(kong)(kong)內液(ye)(ye)體(ti)(ti)。5. 加酶(mei)(mei)(mei)(mei):在(zai)(zai)(zai)每(mei)個(ge)孔(kong)(kong)(kong)(kong)(kong)中(zhong)(zhong)加入酶(mei)(mei)(mei)(mei)標(biao)物(wu)(wu)(wu)(wu)100μL。6. 溫(wen)育(yu):再次將(jiang)酶(mei)(mei)(mei)(mei)標(biao)板(ban)密封(feng),在(zai)(zai)(zai)37℃下(xia)溫(wen)育(yu)30分(fen)(fen)鐘。7. 洗滌(di)(di):用(yong)洗滌(di)(di)液(ye)(ye)清(qing)(qing)洗酶(mei)(mei)(mei)(mei)標(biao)板(ban),共洗滌(di)(di)5次。每(mei)次洗滌(di)(di)后(hou),用(yong)吸(xi)水(shui)紙輕輕吸(xi)干孔(kong)(kong)(kong)(kong)(kong)內液(ye)(ye)體(ti)(ti)。8. 加底(di)物(wu)(wu)(wu)(wu):在(zai)(zai)(zai)每(mei)個(ge)孔(kong)(kong)(kong)(kong)(kong)中(zhong)(zhong)加入底(di)物(wu)(wu)(wu)(wu)A液(ye)(ye)50μL,再加入底(di)物(wu)(wu)(wu)(wu)B液(ye)(ye)50μL。9. 終止反應:在(zai)(zai)(zai)每(mei)個(ge)孔(kong)(kong)(kong)(kong)(kong)中(zhong)(zhong)加入終止液(ye)(ye)50μL。10. 檢測(ce):用(yong)酶(mei)(mei)(mei)(mei)標(biao)儀在(zai)(zai)(zai)450nm波長下(xia)讀(du)(du)取(qu)各孔(kong)(kong)(kong)(kong)(kong)的(de)(de)(de)吸(xi)光(guang)度(OD值)。 結果(guo)分(fen)(fen)析根(gen)據標(biao)準(zhun)(zhun)品和(he)樣(yang)本的(de)(de)(de)OD值,繪制標(biao)準(zhun)(zhun)曲(qu)線,從而計算出(chu)樣(yang)本中(zhong)(zhong)角蛋白(bai)1(KRT1)的(de)(de)(de)濃度。具體(ti)(ti)分(fen)(fen)析方法可參考(kao)試(shi)劑(ji)盒說明書(shu)(shu)或相(xiang)關文獻。注意事項(xiang)1. 在(zai)(zai)(zai)操作過程中(zhong)(zhong),應保持(chi)酶(mei)(mei)(mei)(mei)標(biao)板(ban)的(de)(de)(de)干燥,避(bi)免(mian)水(shui)分(fen)(fen)進(jin)入孔(kong)(kong)(kong)(kong)(kong)內。2. 加樣(yang)時(shi)需(xu)(xu)(xu)確(que)保加樣(yang)準(zhun)(zhun)確(que)、無(wu)氣泡產生。3. 溫(wen)育(yu)和(he)洗滌(di)(di)過程中(zhong)(zhong)需(xu)(xu)(xu)嚴格控制時(shi)間和(he)溫(wen)度,確(que)保實驗結果(guo)的(de)(de)(de)準(zhun)(zhun)確(que)性(xing)。4. 在(zai)(zai)(zai)使(shi)用(yong)酶(mei)(mei)(mei)(mei)標(biao)儀讀(du)(du)取(qu)OD值時(shi),需(xu)(xu)(xu)確(que)保波長設置(zhi)正(zheng)確(que),避(bi)免(mian)誤差產生。5. 在(zai)(zai)(zai)整個(ge)實驗過程中(zhong)(zhong),需(xu)(xu)(xu)保持(chi)操作環境的(de)(de)(de)清(qing)(qing)潔(jie)和(he)整潔(jie),避(bi)免(mian)交叉污染(ran)。
猴粘蛋(dan)白17(MUC17)酶聯免(mian)(mian)疫試劑(ji)(ji)盒實驗(yan)原(yuan)(yuan)理 ELISA運(yun)用(yong)了免(mian)(mian)疫學(xue)原(yuan)(yuan)理和化學(xue)反(fan)應顯色(se)原(yuan)(yuan)理,需要(yao)將抗原(yuan)(yuan)或抗體(ti)預(yu)包被在固相載(zai)(zai)體(ti)表(biao)面(mian),使(shi)后(hou)來形成(cheng)(cheng)的(de)(de)(de)抗原(yuan)(yuan)-抗體(ti)-酶 (熒(ying)光(guang)基團) -底(di)物(wu)(wu)(wu)復合物(wu)(wu)(wu)黏附在載(zai)(zai)體(ti)上,通(tong)過檢測(ce)OD值(zhi)或發光(guang)值(zhi),來檢測(ce)靶蛋(dan)白的(de)(de)(de)含量。 實驗(yan)所需自(zi)備物(wu)(wu)(wu)品(pin)1. 酶標儀(yi)(450nm)2. 加樣(yang)器(qi)及槍頭:0.5-10uL、2-20uL、20-200uL、200-1000uL3. 37℃恒溫(wen)(wen)箱(xiang) 試劑(ji)(ji)盒組成(cheng)(cheng)名稱96 孔(kong)(kong)(kong)配置(zhi)48 孔(kong)(kong)(kong)配置(zhi)備注微孔(kong)(kong)(kong)酶標板(ban)(ban)12 孔(kong)(kong)(kong)×8 條12 孔(kong)(kong)(kong)×4 條無(wu)(wu)標準(zhun)品(pin)0.5mL0.5mL按說(shuo)明(ming)(ming)書(shu)進(jin)(jin)(jin)(jin)(jin)行稀釋(shi)(shi)標準(zhun)品(pin)稀釋(shi)(shi)液(ye)6mL3mL按說(shuo)明(ming)(ming)書(shu)進(jin)(jin)(jin)(jin)(jin)行稀釋(shi)(shi)樣(yang)本稀釋(shi)(shi)液(ye)6mL3mL按說(shuo)明(ming)(ming)書(shu)進(jin)(jin)(jin)(jin)(jin)行稀釋(shi)(shi)檢測(ce)抗體(ti)-HRP6mL3mL無(wu)(wu)20×洗(xi)(xi)(xi)滌(di)(di)緩(huan)沖液(ye)20mL20mL按說(shuo)明(ming)(ming)書(shu)進(jin)(jin)(jin)(jin)(jin)行稀釋(shi)(shi)底(di)物(wu)(wu)(wu) A6mL3mL無(wu)(wu)底(di)物(wu)(wu)(wu) B6mL3mL無(wu)(wu)終止液(ye)6mL3mL無(wu)(wu)封(feng)板(ban)(ban)膜2 張2 張無(wu)(wu)說(shuo)明(ming)(ming)書(shu)1 份1 份無(wu)(wu)自(zi)封(feng)袋1 個(ge)1 個(ge)無(wu)(wu) 操作步(bu)驟(zou)1. 準(zhun)備工作:將試劑(ji)(ji)盒從(cong)冰箱(xiang)中(zhong)(zhong)取(qu)出(chu),室溫(wen)(wen)放置(zhi)30分鐘(zhong),使(shi)試劑(ji)(ji)盒恢復至室溫(wen)(wen)。同時(shi)準(zhun)備好實驗(yan)所需的(de)(de)(de)材(cai)料和器(qi)具。2. 加樣(yang):在酶標板(ban)(ban)的(de)(de)(de)弟(di)1個(ge)孔(kong)(kong)(kong)中(zhong)(zhong)加入(ru)標準(zhun)品(pin)50μL,然后(hou)在后(hou)續的(de)(de)(de)孔(kong)(kong)(kong)中(zhong)(zhong)依次加入(ru)待測(ce)樣(yang)本50μL。3. 溫(wen)(wen)育:將酶標板(ban)(ban)密封(feng),在37℃下溫(wen)(wen)育30分鐘(zhong)。4. 洗(xi)(xi)(xi)滌(di)(di):用(yong)洗(xi)(xi)(xi)滌(di)(di)液(ye)清洗(xi)(xi)(xi)酶標板(ban)(ban),共(gong)洗(xi)(xi)(xi)滌(di)(di)5次。每(mei)(mei)次洗(xi)(xi)(xi)滌(di)(di)后(hou),用(yong)吸(xi)水(shui)紙(zhi)(zhi)輕(qing)(qing)輕(qing)(qing)吸(xi)干孔(kong)(kong)(kong)內(nei)(nei)液(ye)體(ti)。5. 加酶:在每(mei)(mei)個(ge)孔(kong)(kong)(kong)中(zhong)(zhong)加入(ru)酶標物(wu)(wu)(wu)100μL。6. 溫(wen)(wen)育:再次將酶標板(ban)(ban)密封(feng),在37℃下溫(wen)(wen)育30分鐘(zhong)。7. 洗(xi)(xi)(xi)滌(di)(di):用(yong)洗(xi)(xi)(xi)滌(di)(di)液(ye)清洗(xi)(xi)(xi)酶標板(ban)(ban),共(gong)洗(xi)(xi)(xi)滌(di)(di)5次。每(mei)(mei)次洗(xi)(xi)(xi)滌(di)(di)后(hou),用(yong)吸(xi)水(shui)紙(zhi)(zhi)輕(qing)(qing)輕(qing)(qing)吸(xi)干孔(kong)(kong)(kong)內(nei)(nei)液(ye)體(ti)。8. 加底(di)物(wu)(wu)(wu):在每(mei)(mei)個(ge)孔(kong)(kong)(kong)中(zhong)(zhong)加入(ru)底(di)物(wu)(wu)(wu)A液(ye)50μL,再加入(ru)底(di)物(wu)(wu)(wu)B液(ye)50μL。9. 終止反(fan)應:在每(mei)(mei)個(ge)孔(kong)(kong)(kong)中(zhong)(zhong)加入(ru)終止液(ye)50μL。10. 檢測(ce):用(yong)酶標儀(yi)在450nm波長下讀(du)取(qu)各孔(kong)(kong)(kong)的(de)(de)(de)吸(xi)光(guang)度(OD值(zhi))。 結果分析根據標準(zhun)品(pin)和樣(yang)本的(de)(de)(de)OD值(zhi),繪(hui)制(zhi)標準(zhun)曲線,從(cong)而計算出(chu)樣(yang)本中(zhong)(zhong)角蛋(dan)白1(KRT1)的(de)(de)(de)濃度。具體(ti)分析方法可參考試劑(ji)(ji)盒說(shuo)明(ming)(ming)書(shu)或相關(guan)文獻。注意(yi)事項1. 在操作過程中(zhong)(zhong),應保(bao)持(chi)酶標板(ban)(ban)的(de)(de)(de)干燥,避(bi)免(mian)(mian)水(shui)分進(jin)(jin)(jin)(jin)(jin)入(ru)孔(kong)(kong)(kong)內(nei)(nei)。2. 加樣(yang)時(shi)需確(que)(que)(que)保(bao)加樣(yang)準(zhun)確(que)(que)(que)、無(wu)(wu)氣泡產生。3. 溫(wen)(wen)育和洗(xi)(xi)(xi)滌(di)(di)過程中(zhong)(zhong)需嚴格(ge)控制(zhi)時(shi)間和溫(wen)(wen)度,確(que)(que)(que)保(bao)實驗(yan)結果的(de)(de)(de)準(zhun)確(que)(que)(que)性。4. 在使(shi)用(yong)酶標儀(yi)讀(du)取(qu)OD值(zhi)時(shi),需確(que)(que)(que)保(bao)波長設置(zhi)正確(que)(que)(que),避(bi)免(mian)(mian)誤差產生。5. 在整個(ge)實驗(yan)過程中(zhong)(zhong),需保(bao)持(chi)操作環境的(de)(de)(de)清潔和整潔,避(bi)免(mian)(mian)交(jiao)叉污染。
猴突觸核蛋白γ(SNCγ)酶(mei)(mei)聯免(mian)疫試劑(ji)(ji)盒(he) 實(shi)(shi)驗(yan)原(yuan)(yuan)理 ELISA運用(yong)(yong)(yong)了免(mian)疫學(xue)原(yuan)(yuan)理和化學(xue)反應顯色(se)原(yuan)(yuan)理,需(xu)(xu)要(yao)將抗原(yuan)(yuan)或抗體(ti)(ti)(ti)預包被在(zai)(zai)(zai)(zai)固(gu)相(xiang)載體(ti)(ti)(ti)表面(mian),使后來(lai)形成的抗原(yuan)(yuan)-抗體(ti)(ti)(ti)-酶(mei)(mei) (熒光基團) -底(di)(di)物(wu)(wu)(wu)復合物(wu)(wu)(wu)黏附在(zai)(zai)(zai)(zai)載體(ti)(ti)(ti)上(shang),通過(guo)檢(jian)測(ce)(ce)OD值(zhi)或發光值(zhi),來(lai)檢(jian)測(ce)(ce)靶蛋白的含量(liang)。 實(shi)(shi)驗(yan)所(suo)需(xu)(xu)自備物(wu)(wu)(wu)品(pin)1. 酶(mei)(mei)標(biao)(biao)(biao)儀(yi)(yi)(450nm)2. 加(jia)(jia)(jia)樣(yang)(yang)(yang)器及槍(qiang)頭(tou):0.5-10uL、2-20uL、20-200uL、200-1000uL3. 37℃恒溫(wen)(wen)箱(xiang)(xiang) 試劑(ji)(ji)盒(he)組成名(ming)稱(cheng)96 孔(kong)配(pei)置(zhi)48 孔(kong)配(pei)置(zhi)備注微孔(kong)酶(mei)(mei)標(biao)(biao)(biao)板12 孔(kong)×8 條12 孔(kong)×4 條無(wu)標(biao)(biao)(biao)準(zhun)品(pin)0.5mL0.5mL按(an)說(shuo)明(ming)書(shu)(shu)(shu)進(jin)行(xing)稀(xi)釋標(biao)(biao)(biao)準(zhun)品(pin)稀(xi)釋液(ye)(ye)(ye)6mL3mL按(an)說(shuo)明(ming)書(shu)(shu)(shu)進(jin)行(xing)稀(xi)釋樣(yang)(yang)(yang)本稀(xi)釋液(ye)(ye)(ye)6mL3mL按(an)說(shuo)明(ming)書(shu)(shu)(shu)進(jin)行(xing)稀(xi)釋檢(jian)測(ce)(ce)抗體(ti)(ti)(ti)-HRP6mL3mL無(wu)20×洗(xi)(xi)(xi)滌緩沖液(ye)(ye)(ye)20mL20mL按(an)說(shuo)明(ming)書(shu)(shu)(shu)進(jin)行(xing)稀(xi)釋底(di)(di)物(wu)(wu)(wu) A6mL3mL無(wu)底(di)(di)物(wu)(wu)(wu) B6mL3mL無(wu)終止(zhi)液(ye)(ye)(ye)6mL3mL無(wu)封(feng)板膜2 張2 張無(wu)說(shuo)明(ming)書(shu)(shu)(shu)1 份1 份無(wu)自封(feng)袋1 個(ge)(ge)(ge)1 個(ge)(ge)(ge)無(wu) 操(cao)作步驟1. 準(zhun)備工作:將試劑(ji)(ji)盒(he)從冰箱(xiang)(xiang)中取(qu)出,室溫(wen)(wen)放(fang)置(zhi)30分鐘,使試劑(ji)(ji)盒(he)恢復至室溫(wen)(wen)。同(tong)時(shi)(shi)準(zhun)備好實(shi)(shi)驗(yan)所(suo)需(xu)(xu)的材(cai)料(liao)和器具。2. 加(jia)(jia)(jia)樣(yang)(yang)(yang):在(zai)(zai)(zai)(zai)酶(mei)(mei)標(biao)(biao)(biao)板的弟(di)1個(ge)(ge)(ge)孔(kong)中加(jia)(jia)(jia)入標(biao)(biao)(biao)準(zhun)品(pin)50μL,然后在(zai)(zai)(zai)(zai)后續(xu)的孔(kong)中依次(ci)加(jia)(jia)(jia)入待測(ce)(ce)樣(yang)(yang)(yang)本50μL。3. 溫(wen)(wen)育(yu):將酶(mei)(mei)標(biao)(biao)(biao)板密(mi)(mi)封(feng),在(zai)(zai)(zai)(zai)37℃下溫(wen)(wen)育(yu)30分鐘。4. 洗(xi)(xi)(xi)滌:用(yong)(yong)(yong)洗(xi)(xi)(xi)滌液(ye)(ye)(ye)清洗(xi)(xi)(xi)酶(mei)(mei)標(biao)(biao)(biao)板,共洗(xi)(xi)(xi)滌5次(ci)。每次(ci)洗(xi)(xi)(xi)滌后,用(yong)(yong)(yong)吸(xi)(xi)水(shui)紙輕輕吸(xi)(xi)干孔(kong)內(nei)液(ye)(ye)(ye)體(ti)(ti)(ti)。5. 加(jia)(jia)(jia)酶(mei)(mei):在(zai)(zai)(zai)(zai)每個(ge)(ge)(ge)孔(kong)中加(jia)(jia)(jia)入酶(mei)(mei)標(biao)(biao)(biao)物(wu)(wu)(wu)100μL。6. 溫(wen)(wen)育(yu):再(zai)次(ci)將酶(mei)(mei)標(biao)(biao)(biao)板密(mi)(mi)封(feng),在(zai)(zai)(zai)(zai)37℃下溫(wen)(wen)育(yu)30分鐘。7. 洗(xi)(xi)(xi)滌:用(yong)(yong)(yong)洗(xi)(xi)(xi)滌液(ye)(ye)(ye)清洗(xi)(xi)(xi)酶(mei)(mei)標(biao)(biao)(biao)板,共洗(xi)(xi)(xi)滌5次(ci)。每次(ci)洗(xi)(xi)(xi)滌后,用(yong)(yong)(yong)吸(xi)(xi)水(shui)紙輕輕吸(xi)(xi)干孔(kong)內(nei)液(ye)(ye)(ye)體(ti)(ti)(ti)。8. 加(jia)(jia)(jia)底(di)(di)物(wu)(wu)(wu):在(zai)(zai)(zai)(zai)每個(ge)(ge)(ge)孔(kong)中加(jia)(jia)(jia)入底(di)(di)物(wu)(wu)(wu)A液(ye)(ye)(ye)50μL,再(zai)加(jia)(jia)(jia)入底(di)(di)物(wu)(wu)(wu)B液(ye)(ye)(ye)50μL。9. 終止(zhi)反應:在(zai)(zai)(zai)(zai)每個(ge)(ge)(ge)孔(kong)中加(jia)(jia)(jia)入終止(zhi)液(ye)(ye)(ye)50μL。10. 檢(jian)測(ce)(ce):用(yong)(yong)(yong)酶(mei)(mei)標(biao)(biao)(biao)儀(yi)(yi)在(zai)(zai)(zai)(zai)450nm波(bo)長(chang)(chang)下讀取(qu)各孔(kong)的吸(xi)(xi)光度(du)(OD值(zhi))。 結果分析(xi)根(gen)據標(biao)(biao)(biao)準(zhun)品(pin)和樣(yang)(yang)(yang)本的OD值(zhi),繪制標(biao)(biao)(biao)準(zhun)曲線,從而計算出樣(yang)(yang)(yang)本中角蛋白1(KRT1)的濃度(du)。具體(ti)(ti)(ti)分析(xi)方(fang)法可參考試劑(ji)(ji)盒(he)說(shuo)明(ming)書(shu)(shu)(shu)或相(xiang)關(guan)文獻(xian)。注意事(shi)項1. 在(zai)(zai)(zai)(zai)操(cao)作過(guo)程(cheng)中,應保(bao)持酶(mei)(mei)標(biao)(biao)(biao)板的干燥(zao),避免(mian)水(shui)分進(jin)入孔(kong)內(nei)。2. 加(jia)(jia)(jia)樣(yang)(yang)(yang)時(shi)(shi)需(xu)(xu)確保(bao)加(jia)(jia)(jia)樣(yang)(yang)(yang)準(zhun)確、無(wu)氣泡產生。3. 溫(wen)(wen)育(yu)和洗(xi)(xi)(xi)滌過(guo)程(cheng)中需(xu)(xu)嚴(yan)格控制時(shi)(shi)間和溫(wen)(wen)度(du),確保(bao)實(shi)(shi)驗(yan)結果的準(zhun)確性。4. 在(zai)(zai)(zai)(zai)使用(yong)(yong)(yong)酶(mei)(mei)標(biao)(biao)(biao)儀(yi)(yi)讀取(qu)OD值(zhi)時(shi)(shi),需(xu)(xu)確保(bao)波(bo)長(chang)(chang)設(she)置(zhi)正(zheng)確,避免(mian)誤差產生。5. 在(zai)(zai)(zai)(zai)整個(ge)(ge)(ge)實(shi)(shi)驗(yan)過(guo)程(cheng)中,需(xu)(xu)保(bao)持操(cao)作環境的清潔和整潔,避免(mian)交叉污染(ran)。