BCA蛋白浓度测定试剂盒
产品编号
产品名称
包装
P0012 BCA蛋白浓度测定试剂盒 500次
产品简介：
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BCA蛋白浓度测定试剂盒(BCA Protein Assay Kit)是根据目前世界上最常用的两种蛋白浓度检测方法之一BCA法研制而 成，实现了蛋白浓度测定的简单，高稳定性，高灵敏度和高兼容性。
灵敏度高，检测浓度下限达到25µg/ml，最小检测蛋白量达到0.5µg，待测样品体积为1-20µl。 在50-2000µg/ml浓度范围内有较好的线性关系。
BCA法测定蛋白浓度不受绝大部分样品中的化学物质的影响，可以兼容样品中高达5%的SDS，5%的Triton X-100，5%的Tween 20, 60, 80。但受螯合剂和略高浓度的还原剂的影响，需确保EDTA低于10mM，无EGTA，二硫苏糖醇低于1mM，β-巯基乙醇低于0.01%。不适用BCA法时建议试用碧云天生产的Bradford蛋白浓度测定试剂盒(P0006)。 BCA蛋白浓度测定试剂盒对样品中各种物质的详细的兼容性，请参见碧云天如下网页: http://www.beyotime.com/Compatibility Chart For BCA Kit.pdf 每个试剂盒可以检测500个样品。
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包装清单：
产品编号 产品名称 P0012-1 BCA试剂 A P0012-2 BCA试剂 B P0012-3 蛋白标准(5mg/ml BSA) — 说明书
包装
100ml 3ml 1ml 1份
保存条件：
BCA试剂A和B室温保存，蛋白标准请-20℃冻存。本试剂盒自订购之日起一年内有效。
注意事项：
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需酶标仪一台，测定波长为540-595nm之间，562nm最佳。需96孔板。如果没有酶标仪，也可以使用普通的分光光度计测定，但测定时，需根据比色皿的最小检测体积，适当加大BCA工作液的用量使不小于最小检测体积，样品和标准品的用量可相应按比例放大也可不变。使用分光光度计测定蛋白浓度时，每个试剂盒可以测定的样品数量可能会显著减少。 如发现样品稀释液或裂解液本身就有较高背景，请试用碧云天生产的Bradford蛋白浓度测定试剂盒(P0006)。 为了加快BCA法测定蛋白浓度的速度可以适当用微波炉加热，但是切勿过热。
EDTA浓度必须小于10mM，不兼容EGTA。不适用BCA法时，请试用碧云天生产的Bradford蛋白浓度测定试剂盒(P0006)。 为了您的安全和健康，请穿实验服并戴一次性手套操作。
根据样品数量，按50体积BCA试剂A加1体积BCA试剂B(50:1)配制适量BCA工作液，充分混匀。BCA工作液室温24小时内稳定。
完全溶解蛋白标准品，取10µl稀释至100µl，使终浓度为0.5mg/ml。蛋白样品在什么溶液中，标准品也宜用什么溶液稀释。但是为了简便起见，也可以用0.9%NaCl或PBS稀释标准品。
将标准品按0, 1, 2, 4, 8, 12, 16, 20µl加到96孔板的标准品孔中，加用于稀释标准品的溶液补足到20µl。 加适当体积样品到96孔板的样品孔中，加用于稀释标准品的溶液到20µl。 各孔加入200µl BCA工作液，37℃放置30分钟。
注: 也可以室温放置2小时，或60℃放置30分钟。BCA法测定蛋白浓度时，吸光度会随着时间的延长不断加深。并且显色反应会因温度升高而加快。如果浓度较低，适合在较高温度孵育，或延长孵育时间。 测定A562，540-595nm之间的波长也可接受。根据标准曲线计算出蛋白浓度。
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使用说明：
6.
常见问题：
1. 测定标准曲线时发现随着标准品浓度的增加吸光度或颜色没有明显变化。
可能的原因是样品中含有严重干扰BCA法测定蛋白浓度的物质，详细的BCA法的兼容性列表请参考碧云天如下网页: http://www.beyotime.com/Compatibility Chart For BCA Kit.pdf 2. 是否每次测定时都需要做标准曲线？
建议每次测定时都做标准曲线。因为BCA法测定时颜色会随着时间的延长不断加深，并且显色反应的速度和温度有关，所以除非精确控制显色反应的时间和温度，否则如需精确测定宜每次都做标准曲线。 使用本产品的文献：
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