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- 文献和实验
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- 库存:
大量
- 供应商:
上海钰博生物科技有限公司
- 检测范围:
0.312-20ng/mL
- 检测方法:
用于科研实验检测,定量定性检测
- 应用:
科研使用
- 适应物种:
Homo sapiens human
- 标记物:
血清,血浆,尿液及相关液体等
- 样本:
人,大鼠,小鼠,兔,猪,植物等
- 规格:
48T/96T
英文名称:Human Plasmin/Antiplasmin Complex (PAP) ELISA Kit
检测范围:0.312-20ng/mL
灵敏度:0.144ng/mL
规格:48T盒/96T盒
贮藏:2-8℃,避光防潮保存,有效期:6个月
用途:本产品仅供科研,不能用于临床诊断
PAP试剂盒描述:
酶联免疫吸附试验(ELISA)用于检测样品中抗原的存在。抗原通过吸附作用固定在板的阱中,或者用结合的抗原特异性抗体捕获。然后,如果存在抗体,则加入检测抗体与抗原形成复合物。检测抗体可以共价连接到一种酶,或者由二级酶联抗体检测。酶底物然后加入到威尔斯产生一个可见的信号,这是与抗原的数量,并用分光光度计测量。
PAP试剂盒程序:
一般ELISA仅供参考。ELISA和EIA试剂盒具体协议的问题,请参照试剂盒说明,100µL肽(@ 4µ克/毫升)涂层的缓冲区添加到微量滴定板的个人威尔斯。培养板2小时37°C或隔夜在4°C.除去包衣液洗板三次,100μL pbs-0.05 % Tween20填充威尔斯。该解决方案或水洗是轻弹板在厨房里删除。剩下的水滴通过在纸上拍盘子而被去除。
在包裹的威尔斯中,用100微米的阻断缓冲液,3%的脱脂乳在PBS中堵塞其余的蛋白质结合位点。培养1小时室温轻轻摇动。洗板三次,以100ul pbs-0.05吐温20。添加50µl稀释的抗体在每。培养板在37℃为轻轻摇动一个小时。洗板六次,以100ul pbs-0.05吐温20。使用前加入50克共轭二次抗体,按最佳浓度稀释。就在37℃一小时。洗板六次,以100ul pbs-0.05 % Tween20。通过混合乙酸制备的底物溶液,TMB和0.03%的H2O2与1:4:5体积比。
PAP试剂盒用多通道管将每个溶液的50微米溶液分配给每个井。培养板在黑暗中37℃15-30分钟。
在足够的颜色显影后,在威尔斯(如果需要的话)中添加100微米的停止液。
读取吸光度(在450nm处的光密度)每个在读板。
PAP试剂盒的基本原理有三条:
(1)抗原或抗体能以物理性地吸附于固相载体表面,可能是蛋白和聚苯乙烯表面间的疏水性部分相互吸附,并保持其免疫学活性。
(2)抗原或抗体可通过共价键与酶连接形成酶结合物,而此种酶结合物仍能保持其免疫学和酶学活性。
(3)酶结合物与相应抗原或抗体结合后,可根据加入底物的颜色反应来判定是否有免疫反应的存在,而且颜色反应的深浅是与标本中相应抗原或抗体的量成正比例的,因此,可以按底物显色的程度显示试验结果。
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