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人(ren)(Human)胰(yi)蛋(dan)白(bai)酶(mei)原(yuan)Ⅱ(Try-Ⅱ)ELISA檢(jian)測(ce)試劑(ji)盒
發(fa)布(bu)時(shi)間(jian):2012/4/12
點擊(ji)次數:499
本試劑(ji)盒只(zhi)能用(yong)於(yu)科(ke)學研(yan)究(jiu),不得(de)用於(yu)醫(yi)學診斷(duan)
人(Human)胰(yi)蛋(dan)白(bai)酶(mei)原(yuan)Ⅱ(Try-Ⅱ)ELISA檢(jian)測(ce)試劑(ji)盒
使(shi)用(yong)說(shuo)明(ming)書(shu)
檢(jian)測(ce)原(yuan)理
試劑(ji)盒采(cai)用雙抗(kang)體壹(yi)步(bu)夾(jia)心法(fa)酶(mei)聯免疫(yi)吸附試驗(yan)(ELISA)。往預
先(xian)包被胰(yi)蛋(dan)白(bai)酶(mei)原(yuan)Ⅱ(Try-Ⅱ)抗(kang)體的(de)包(bao)被微(wei)孔中(zhong),依(yi)次加(jia)入標(biao)本、
標(biao)準(zhun)品(pin)、HRP標(biao)記(ji)的(de)檢(jian)測(ce)抗(kang)體,經(jing)過溫育並(bing)*洗滌(di)。用(yong)底物TMB顯
色,TMB在(zai)過氧化物酶(mei)的(de)催化(hua)下(xia)轉(zhuan)化(hua)成藍(lan)色(se),並(bing)在(zai)酸(suan)的作(zuo)用下(xia)轉(zhuan)化(hua)成
zui終的黃(huang)色(se)。顏(yan)色(se)的(de)深淺(qian)和(he)樣品中(zhong)的(de)胰(yi)蛋(dan)白(bai)酶(mei)原(yuan)Ⅱ(Try-Ⅱ)呈(cheng)正(zheng)相(xiang)
關。用(yong)酶(mei)標(biao)儀(yi)在(zai)450nm 波長下(xia)測(ce)定(ding)吸光度(du)(OD 值),計(ji)算(suan)樣品濃(nong)度(du)。
樣品收集(ji)、處(chu)理及(ji)保存(cun)方(fang)法(fa)
1. 血清(qing):使(shi)用(yong)不含(han)熱(re)原(yuan)和(he)內毒(du)素的(de)試管(guan),操作過程中(zhong)避(bi)免(mian)任(ren)何(he)細胞(bao)
刺(ci)激,收集(ji)血(xue)液後(hou),3000 轉(zhuan)離心10 分鐘(zhong)將血清(qing)和紅細胞(bao)迅(xun)速(su)小(xiao)心(xin)地(di)
分離。
2. 血漿:EDTA、檸(ning)檬酸(suan)鹽或(huo)肝(gan)素抗(kang)凝(ning)。3000 轉(zhuan)離(li)心(xin)30 分鐘(zhong)取上(shang)清(qing)。
3. 細胞(bao)上(shang)清(qing)液:3000 轉離心(xin)10 分鐘(zhong)去(qu)除顆粒和(he)聚(ju)合物。
4. 組織勻(yun)漿(jiang):將組織加(jia)入適量生理鹽水(shui)搗(dao)碎(sui)。3000 轉離(li)心10 分(fen)鐘(zhong)
取上(shang)清(qing)。
5. 保存(cun):如果樣本收集(ji)後(hou)不(bu)及時檢測(ce),請按(an)壹(yi)次(ci)用量分裝,凍(dong)存於(yu)
-20℃,避(bi)免(mian)反復凍融,在(zai)室(shi)溫下(xia)解(jie)凍(dong)並(bing)確保樣品均勻(yun)地(di)充(chong)分(fen)解(jie)凍(dong)。
自(zi)備物品(pin)
1. 酶(mei)標(biao)儀(yi)(450nm)
2. 高(gao)精(jing)度(du)加樣器及槍(qiang)頭(tou):0.5-10uL、2-20uL、20-200uL、200-1000uL
3. 37℃恒溫箱
操作註意事(shi)項(xiang)
1. 試劑(ji)盒保存(cun)在(zai)2-8℃,使(shi)用(yong)前(qian)室溫平衡(heng)20 分(fen)鐘(zhong)。從(cong)冰(bing)箱取出的
濃(nong)縮洗滌液會(hui)有(you)結晶(jing),這(zhe)屬於(yu)正(zheng)常現(xian)象,水(shui)浴(yu)加熱(re)使(shi)結晶(jing)*溶解(jie)
後(hou)再(zai)使(shi)用(yong)。
2. 實(shi)驗(yan)中(zhong)不(bu)用(yong)的(de)板條應(ying)立(li)即(ji)放(fang)回(hui)自(zi)封(feng)袋中(zhong),密封(feng)(低溫幹燥(zao))保存(cun)。
3. 濃度(du)為0 的(de)S0 號標(biao)準(zhun)品(pin)即(ji)可(ke)視(shi)為(wei)陰性對(dui)照或(huo)者(zhe)空(kong)白;按(an)照(zhao)說(shuo)明(ming)obtained for each of the six standard concentrations on the vertical (Y) axis
versus the corresponding concentration on the horizontal (X) axis.
2. First, calculate the mean O.D. value for each standard and sample. All O.D.
values, are subtracted by the mean value of the zero standard before result
interpretation. Construct the standard curve using graph paper or statistical
software.
3. To determine the amount in each sample, first locate the O.D. value on the
Y-axis and extend a horizontal line to the standard curve. At the point of
intersection, draw a vertical line to the X-axis and read the corresponding
concentration.
4. Any variation in operator, pipetting and washing technique, incubation time or
temperature, and kit age can cause variation in result. Each user should obtain
their own standard curve.
5. The sensitivity by this assay is 1.0 ng/ml
6. Standard curve
Storage: 2-8℃.
validity: six months.
FOR RESEARCH USE ONLY; NOT FOR THERAPEUTIC OR
DIAGNOSTICAPPLICATIONS! PLEASE READ THROUGH
ENTIRE PROCEDURE BEFORE BEGINNING!