人环加氧酶1(PTGS1) ELISA试剂盒常见问题解析：
1.问：我的标准曲线看起来很好，但我没有在样本中得到预期的信号，这是为什么？答：样品可能不含分析物。矩阵效应可能掩盖检测。确保按照我司试剂盒中说明书进行稀释。稀释，前请检查以确保稀释操作正确。过度稀释可能导致样品低于标准曲线的范围。Q:My standard curve looked fine, but I didn’t get a signal in my sample when I expected to, why?A:The sample may not contain the analyte. A matrix effect may be masking the detection. Ensure that the recommended dilution was followed as stated in the kit insert. If dilution was recommended, check to be sure that the dilution was performed properly. Over-dilution may cause the sample to fall below the range of the standard curve.
2.问：你建议我该如何洗板？答：如果您使用的是自动洗板机，我们建议定期检查校正，并在清洗前用洗板机缓冲液冲洗系统。手动洗板机也是如此。或者使用中继器或洗涤瓶。用户应小心确保所有内容物都被吸入，并在无棉纸上轻敲干燥。Q:How do you recommend I wash my plate?A:If you are using an automated plate washer we recommend that the calibration be checked on a regular basis, and that the system is flushed with the Plate Washing Buffer prior to washing. The same is true for a manual washer. A repeater or a wash bottle can also be used. The user should be careful to ensure that all of the contents are aspirated and the plate tapped dry on lint-free paper.
【血浆样本】: 避免使用溶血样品抗凝剂推荐使用EDTA-Na2,样品采集后30分钟内于1000×g离心15分钟,取上清即可检测。
【组织匀浆】:用预冷的PBS (0.01M, pH=7.4)冲洗组织,去除残留血液(匀浆中裂解的红细胞会影响测量结果),称重后将组织剪碎。将剪碎的组织与对应体积的PBS(一般按1:9的重量体积比,比如1g的组织样品对应9mL的PBS,具体体积可根据实验需要适当调整,并做好记录。推荐在PBS中加入蛋白酶抑制剂)加入玻璃匀浆器中,于冰上充分研磨。为了进一步裂解组织细胞,可以对匀浆液进行超声破碎,或反复冻融。 将匀浆液于5000×g离心5~10分钟,取上清检测。
【细胞培养上清】:取细胞培养上清于1000×g离心20分钟,除去杂质及细胞碎片。取上清检测。
注意事项：样品应清澈透明,悬浮物应离心去除。 样品收集后若在1周内进行检测的可保存于4℃,若不能及时检测,请按一次使用量分装,冻存于-20℃(1个月内检测),或-80℃(6个月内检测),避免反复冻融。 如果您的样品中检测物浓度高于标准品值,请根据实际情况,做适当倍数稀释(建议先做预实验,以确定稀释倍数)。
