使用说明：
1. 样品测定的准备：(注意：样品裂解需在4℃或冰上操作)
    对于贴壁细胞：
    吸除培养液，按照6孔板每孔加入200微升裂解液的比例(即相当于细胞培养液量2毫升的1/10)加入裂
    解液，裂解细胞。裂解细胞时为了裂解充分，可以使用移液器进行反复吹打或晃动培养板使裂解液
    充分接触并裂解细胞。通常细胞在接触裂解液后会立即裂解。裂解后4℃ 12000g离心5分钟，取上
    清，用于后续的测定。
    对于悬浮细胞：
    用离心管离心沉淀细胞，弃上清，轻轻弹散细胞，按照6孔板每孔的细胞量加入200微升裂解液的比
    例加入裂解液，裂解细胞。裂解细胞时为了裂解充分可以弹击离心管管底或适当Vortex使裂解液充
    分接触并裂解细胞。通常细胞在接触裂解液后会立即裂解。裂解后4℃ 12000g离心5分钟，取上清，
    用于后续的测定。
    对于组织样品：
    按照每20毫克组织加入约100-200微升裂解液的比例加入裂解液，然后用玻璃匀浆器或其它匀浆设备
    进行匀浆。充分匀浆可以确保组织被完全裂解。裂解后4℃ 12000g离心5分钟，取上清，用于后续的
    测定。
2. 标准曲线测定的准备：
    冰浴上溶解待用试剂，把ATP标准溶液用ATP检测裂解液稀释成适当的浓度梯度。具体的浓度需根据
    样品中ATP的浓度而定。初次检测可以检测0.1、1和10微摩尔/升这几个浓度，在后续的实验中，可
    以根据样品中ATP的浓度对标准品的浓度进行适当调节。
3. ATP检测工作液的配制：
    按照每个样品或标准品需100微升ATP检测工作液的比例配制适当量的ATP检测工作液。把待用试剂在
    冰浴上溶解。取适量的ATP检测试剂，按照1:100的比例用ATP检测试剂稀释液稀释ATP检测试剂。例
    如50微升ATP检测试剂可以加入5微升ATP检测试剂稀释液配制成5毫升ATP检测工作液。稀释后的ATP
    检测试剂即为用于后续实验的ATP检测工作液。ATP检测工作液可在冰浴上暂时保存。
4. ATP浓度的测定：
    a. 加100微升ATP检测工作液到检测孔或检测管内。室温放置3-5分钟，以使本底性的ATP全部被消耗
       掉，从而降低本底。可以一次性把10-20个检测孔或检测管分别加上100微升ATP检测工作液，从
       而节省时间。
    b. 在检测孔或检测管内加上20微升样品或标准品，迅速用枪(微量移液器)混匀，至少间隔2秒后，
       用luminometer或液闪仪测定RLU值或CPM。(注：样品的体积可以自行在10-100微升范围内调节。
       如果样品中的ATP浓度比较低则可以加入100微升样品，如果样品中ATP浓度比较高则可以加入较
       小体积的样品，同时标准品也需要使用相同的体积。如果样品中ATP的浓度特别高，可以用ATP
       检测裂解液稀释样品后再测定。本试剂盒在加入100微升标准品时，大致在10nmol/L-10μmol/L
       的浓度范围内有很好的线性关系。)
    c. 根据标准曲线计算出样品中ATP的浓度。
    d. 为了消除样品制备时由于蛋白量的差异而造成的误差，可以用碧云天生产的BCA蛋白浓度测定试
       剂盒(P0009/P0010/P0010S/P0011/P0012/P0012S)测定样品中的蛋白浓度。然后把ATP的浓度换算
       成nmol/mg蛋白的形式。

常见问题：
1. Luminometer和荧光分光光度计有何不同？
    荧光分光光度计检测的样品本身不能发光，样品需要由特定波长的激发光激发，然后才能产生荧光
    并被荧光分光光度计检测。Luminometer，即化学发光检测仪，检测的样品本身可以发光，不需要激
    发光进行激发。也就是说Luminometer是检测化学发光的仪器。有些型号的荧光分光光度计也具有
    luminometer的功能，即也可以检测化学发光。您所使用的荧光分光光度计能否用于化学发光的测定
    请仔细阅读该仪器的说明书。
2. 可以进行荧光素酶报告基因检测的仪器是否就可以用于本试剂盒的ATP检测？
    是。荧光素酶报告基因的检测原理和本ATP检测试剂盒的原理相同，可以用相同的仪器测定，并且可
    以选择相同的测定参数，例如检测前等待时间为2秒，检测时间为10秒等。
3. 多功能酶标仪是否可以用于本试剂盒的ATP检测？
    不一定。如果该多功能酶标仪具有检测化学发光的功能，即具有luminometer的功能，就可以用于本
    试剂盒的检测，否则就不能了。

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