人纤调蛋白（FMOD）ELISA检测试剂盒

库存 ：5633

供应商 ：齐一生物科技（上海）有限公司

检测限 ：详细请见说明书

检测方法 ：ELISA酶联免疫法

应用 ：科研检测

标记物 ：见说明

样本 ：血清，血浆，尿液，胸腹水，脑脊液，细胞培养上清，组织匀浆等

规格 ：48T/96T

人纤调蛋白（FMOD）ELISA检测试剂盒试剂盒规格：96T/48T
本试剂盒库存：现货
试剂盒保存条件：2-8℃
试剂盒使用范围：仅供科研检测,不得用于临床.
试剂盒发货方式：专门快递免费送货上门
产品价格：价格优惠,欢迎来电洽谈！
试剂盒使用说明书:说明书随货发送,您也可以直接联系我公司在线销售人员索取.全国热线：400-991-0197代理多种品牌进口原装、“ELISA试剂盒”.齐一生物科技有限公司作为酶联免疫供应商,凭借精湛的技术,全方位的售前售后免费代测提供技术服务,共同铸就高品质的产品.多年专业酶免服务,质量保证,可免费提供代测服务,一周内出结果.欢迎来电咨询订购！
酶联免疫吸附实验材料,试剂,溶液
1．酶标板,100μltip头,1mlEp管,湿盒
2．包被稀释液（0.05mol/L碳酸钠-碳酸氢钠buffer,pH9.6）
碳酸钠0.15g,碳酸氢钠0.29g,叠氮钠0.02g,加双蒸水至100ml,调至pH9.6
3.封闭液(5%小牛血清/PBS溶液)
小牛血清5ml,1*PBS(pH7.4)95ml
4.洗涤液(PBST,pH7.4)
NaCl0.8g,KH2PO40.02g,Na2HPO4.12H2O0.29g,KCl0.02g,Tween200.05ml,叠氮钠0.01g,加双蒸水至100ml,调至pH7.4
5.样本稀释液(PBS,pH7.4)
NaCl0.8g,KH2PO40.02g,Na2HPO4.12H2O0.29g,KCl0.02g,叠氮钠0.01g,加双蒸水至100ml,调至pH7.4
6.酶标第2抗体(羊抗兔)稀释范围1:5,000-1:100,000
7.底物液(TMB-过氧化氢尿素溶液)
①底物液A:TMB20mg,无水乙醇10ml,加双蒸水至100ml.
②底物液B(0.1mol/L柠檬酸-0.2mol/L磷酸二氢钠缓冲液,pH5.0-5.4)
Na2HPO41.46g,柠檬酸0.933g,0.75%过氧化氢尿素0.64ml,加三蒸水至100ml,调至pH5.0-5.4
③底物A和B按1:1混合即成TMB-过氧化氢尿素溶液.
8.终止液(2mol/LH2SO4溶液)双蒸水200ml,浓硫酸34ml,(缓慢滴加并不断搅拌),加双蒸水至300ml9.0.9%生理盐水
人纤调蛋白（FMOD）ELISA检测试剂盒操作步骤:
1.包被过程(注意设置空白对照,阴性对照):
将所用抗原用包被稀释液稀释到适当浓度(一般所需抗原包被量为每孔20-200μg),每孔抗原加入100μl,置37℃,4h,或4℃,24h;弃去孔中液体.(为避免蒸发,板上应加盖或将板平放在底部有湿纱布的金属湿盒中)
2.封闭酶标反应孔:
5%小牛血清置37℃封闭40min.封闭时将封闭液加满各反应孔,并去除各孔中的气泡,封闭结束后用洗涤液满孔洗涤3遍,每遍3min.
洗涤方法:吸干孔内反应液,将洗涤液注满板孔,放置2min略作摇动,吸干孔内液,倾去液体后在吸水纸上拍干
洗涤次数3次
3.加入待检测样品(建立合适的浓度梯度):
检测时一般采用1:50-1:400的稀释度,应采用较大稀释体积进行,一般保证样品吸取量>20μl.
将稀释好的样品加入酶标反应孔中,每样品至少加双孔,每孔100μl,置于37℃,40-60min.用洗涤液满孔洗涤3遍,每遍3min.
4.加入酶标抗体:
酶标抗体:根据酶结合物提供商提供的参考工作稀释度进行.37℃,30-60min之间.短于30min往往结果不稳定.每孔加100μl.洗涤同前.
5.加入底物液(现用现配):
TMB-过氧化氢尿素溶液,OPD-过氧化氢底物液系统次之.
底物加入量:每孔100μl,置37℃避光放置3-5分钟,加入终止液显色.
6.终止反应:
每孔加入终止液50μl终止反应,于20min内测定实验结果.
7.结果判断:
OPD显色后采用492nm波长,TMB反应产物检测需要450nm波长.检测时一定要首先进行空白孔系统调零.用测定标本孔的吸收值与一组阴性标本测定孔平均值的比值(P/N)表示,当P/N大于2时作为抗体的效价.(数值的大小依具体检测要求而定.)
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