使用说明：
1. 样品的准备：
   a. 血浆、血清或尿液样品制备后可以直接用于MDA测定。
   b. 组织或细胞可以使用PBS或碧云天的Western及IP细胞裂解液(P0013)等裂解液进行匀浆或裂解。对
      于组织，组织重量占匀浆液或裂解液的比例为10%；对于细胞，每100万细胞使用0.1ml裂解液或匀
      浆液。匀浆或裂解后，1600g离心10分钟取上清用于后续测定。匀浆或裂解等样品制备步骤宜在冰
      浴或4℃进行操作。
   c. 对于组织或细胞样品，样品准备完毕后可以用BCA蛋白浓度测定试剂盒(P0009/P0010/P0010S/
      P0011/P0012/P0012S)测定蛋白浓度，以便于后续计算单位蛋白重量组织或细胞内的MDA含量。
   d. 本试剂盒对于样品中的常见化学成分的兼容性参考下表：

2. 试剂盒的准备工作：
   a. TBA储存液的配制：称取适量TBA，用TBA配制液配制成浓度为0.37%的TBA储存液。例如18.5mg
      TBA用5ml TBA配制液配制，最终浓度即为0.37%。TBA配制液需完全溶解后再使用，可以加热到
      70℃以促进溶解。TBA储存液较难溶解，需加热到70℃，并通过剧烈Vortex以促进溶解。配制好的
      TBA储存液室温避光保存，至少3个月内有效。
   b. MDA检测工作液的配制: 根据待测定的样品数(含对照)，参考下表在临检测前新鲜配制适量的
      MDA检测工作液

      注意: MDA检测工作液较难溶解，可以70℃加热，并剧烈Vortex以促进溶解。也可以通过超声处
      理以促进溶解。配制好的MDA检测工作液必须当天使用。
   c. 标准品的稀释：取适量标准品用蒸馏水稀释至1、2、5、10、20、50μM，用于后续制作标准曲
      线。
3. 样品测定:
   a. 在离心管或其它适当容器内加入0.1ml匀浆液、裂解液或PBS等适当溶液作为空白对照，加入0.1ml
      上述不同浓度标准品用于制作标准曲线，加入0.1ml样品用于测定；随后加入0.2ml MDA检测工作
      液。可参考下表设置检测反应体系：

   b. 混匀后，100℃或沸水浴加热15分钟。加热时务必注意避免液体暴沸溅出。如果使用加热块(Heat
      block)进行加热注意用重物压紧离心管盖；如果使用沸水浴，则需使用可把盖子锁死的离心管或
      螺旋盖离心管，或用Parafilm封住离心管口，用针头刺一小孔。最方便和准确的加热方法是使用
      带有热盖并可以加热0.5ml PCR管的PCR仪。
   c. 水浴冷却至室温，1000g室温离心10分钟。取200微升上清加入到96孔板中，随后用酶标仪在532nm
      测定吸光度。如果不方便测定532nm的吸光度，也可以测定530-540nm之间的吸光度。可以设定
      450nm为参考波长进行双波长测定。
   d. MDA含量的计算：对于血浆、血清或尿液等样品可以直接根据标准曲线计算获得MDA的摩尔浓度，
      对于细胞、或组织样品，计算出样品溶液中的MDA含量后，可以通过单位重量的蛋白含量或组织重
      量等来表示最初样品中的MDA含量，例如μmol/mg蛋白或μmol/mg组织。

常见问题：
   1. 没有检测到MDA。
      可能样品中MDA浓度过低，在检测限之下。在检测组织或细胞的MDA时，请注意使用更多的组织或
   细胞。并注意尽量不要稀释样品。

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