人基质重塑关联蛋白5(MXRA5) ELISA试剂盒原理:试剂盒采用双抗体夹心法酶联免疫吸附试验(ELISA)。往预先包被被检测抗体的包被微孔中,依次加入标本、标准品、HRP标记的检测抗体,经过温育并彻底洗涤。用底物TMB显色,TMB在过氧化物酶的催化下转化成蓝色,并在酸的作用下转化成zui终的黄色。颜色的深浅和样品中的被检测指标呈正相关。用酶标仪在450nm 波长下测定吸光度(OD 值),计算样品浓度。常见问题解答:
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.
实验注意事项:
1)严格按照规定的时间和温度进行温育以保证准确结果,所有试剂都必须在使用前达到室温20-25℃,使用后立即冷藏保存试剂。
2)洗板不正确可以导致不准确的结果,在加入底物前确保尽量吸干孔内液体,温育过程中不要让微孔干燥。
3)消除底板残留的液体和手指印,否则影响OD值。
4)底物显色液应呈无色或很浅的颜色,已经变蓝的底物液不能使用。
5)避免试剂盒标本的交叉污染以免造成错误结果。
6)任何反应试剂不能接触漂白溶剂或漂白溶剂所散发的强烈气体,避免影响试剂中的生物活性。
7)平衡至室温后再打开密封袋以防水滴凝聚在冷板条上。