This kit was developed using broadly reactive polyclonal antibodies to the hundreds of different host cell proteins (HCPs) and is intended for use in determining the presence of E. coli HCP impurities in products manufactured by recombinant expression. Our assay validation has determined the LOD for this kit is ~0.2 ng/ml. Please use Sample Diluent, Item No. I028, with these kits; available separately.

描述
该试剂盒是使用针对数百种不同宿主细胞蛋白 (HCP) 的广泛反应性多克隆抗体开发的，旨在用于确定通过重组表达生产的产品中是否存在大肠杆菌 HCP 杂质。 我们的检测验证已确定该试剂盒的 LOD 为 ~0.2 ng/ml。 请在这些套件中使用样品稀释剂，货号 I028； 可单独购买。

Kit Components
Anti-E. coli:HRP Conjugate
E. coli HCP Standards Set, A-F
TBS Wash Concentrate, 20X
Stop Solution
TMB Substrate for ELISA
Anti-E. coli Coated Microtiter Strips

产品英文名称:Cygnus F410 E.coli HCP ELISA Kit

产品中文名称:Cygnus F410 大肠杆菌宿主细胞ELISA检测试剂盒

Cygnus F410 E.coli HCP 大肠杆菌宿主细胞ELISA检测试剂盒 产品介绍:Cygnus F410 E.coli HCP 大肠杆菌宿主细胞ELISA检测试剂盒主要设计用于检测产物重组表达过程中大肠杆菌宿主细胞污染,试剂盒使用的多克隆抗体对数百种HCPs具有反应性。Cygnus F410 E.coli HCP 大肠杆菌宿主细胞ELISA检测试剂盒检测*浓度可达到 0.2ng/mL。ELISA检测相比Western Blot:Western Blots检测过程复杂并需借助程序分析结果。而且,Western Blot本质上是定性分析,不能得到定量结果,检测灵敏性也受到样本体积的限制。Western Blot在下游纯化过程中检测产物HCPs缺乏灵敏度和特异性。而微量滴定板ELISA检测可以克服上述限制。ELISA具备高度灵敏性,可获得半定量结果,是纯化工艺,过程控制,常规质量检测的*选择。

产品使用时一定要做好保护措施:

0.5M硫磺酸有强腐蚀性,要避免接触眼睛、皮肤和衣物。检测前所有试剂都要平衡到室温。

• Cygnus F410 E.coli HCP ELISA Kit 检测原理:

大肠杆菌E. coli检测是一个双位点酶联免疫检测。样本中包含的E. coli HCPs蛋白与标记有抗-E. coli 抗体的辣根过氧化物酶同时反应,反应发生在之前涂有一层吸附性抗-E. coli 蛋白抗体的微量滴定板中进行。免疫反应构成为夹层式结构:固相抗休-HCP-酶标记抗体。反应结束后清洗微量滴定板去除未结合反应物。添加TMB作用底物反应。微量滴定板读数器上测定水解物浓度,水解物浓度与E. coli  HCPs蛋白浓度成比例。用提供的标准液绘制标准曲线,根据读数可以计算出E. coli HCPs蛋白含量。

• Cygnus F410 大肠杆菌宿主细胞ELISA检测试剂盒检测过程:

取出所有试剂平衡到室温→分别移取25μl Cygnus F410 大肠杆菌宿主细胞ELISA检测试剂盒标准液,对照物和样品至待测孔→移取100μl anti-E. coli:HRP(#F411)至每孔→密封滴定板,摇床室温温育90分钟,180rpm→清空小孔,用纸吸干残留液,350μl清洗缓冲液清洗4次→加100μl TMB 作用物(#F005)→室温静置30分钟,不需摇床→加100μl反应终止液(#F006)→分光光度计450/650nm读数

• Cygnus F410 E.coli HCP ELISA Kit 检测结果分析:

Cygnus F410 E.coli HCP ELISA Kit 标准液(0, 1, 3, 12, 40, 100ng/mL)可用于制作标准曲线,标准液浓度相当于免疫反应HCP总浓度。根据测定的小孔中标准液吸光值绘制标准曲线。Cygnus F410 E.coli HCP ELISA Kit 检测结果数据分析可以使用计算机选配曲线,例如点到点,三次样本函数,或四参数逻辑曲线。不要使用线性回归分析修改样品吸光值,这样会出现显著误差。也可以手工绘图,Y轴标注吸光值,X轴标注浓度。然后根据样品吸光值计算出样品中HCP浓度。(操作时每一浓度标准液都要滴定到两个小孔中,以便计算平均分光光度值,样品也需计算平均吸光值)。

Cygnus F410 E.coli HCP ELISA Kit 组成成份:

• Anti- E. coli:HRP(辣根过氧化物酶标记)            1x12mL    F411(Product#)
• Anti- E. coli coated微量滴定板                        12×8孔   F412
• E. coli HCP标准液(0, 1, 3, 12, 40, 100ng/mL)     1mL/瓶  F413
• Stop Solution终止液(0.5M硫磺酸)             1x12mL F006
• TMB (3,3’,5,5’-四甲基联苯氨)                          1x12mL  F005
• Wash Concentrate清洗物(20倍浓缩)                1x50mL    F004

Cygnus相关试剂盒
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