使用说明：
1. 取1.2ml蛋白标准配制液加入到一管蛋白标准(30mg BSA)中，充分溶解后配制成25mg/ml的蛋白标准溶
   液。配制后可立即使用，也可以-20℃长期保存。
2. 取适量25mg/ml蛋白标准，稀释至终浓度为0.5mg/ml。例如取20μl 25mg/ml蛋白标准，加入980μl稀释
   液即可配制成0.5mg/ml蛋白标准。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。稀释后的0.5mg/ml蛋白标准也可以-20℃长期保存。
3. 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。例如5ml
   BCA试剂A加100μlBCA试剂B，混匀，配制成5.1ml BCA工作液。 BCA工作液室温24小时内稳定。
4. 将标准品按0, 1, 2, 4, 8, 12, 16, 20μl加到96孔板的标准品孔中，加标准品稀释液补足到20μl。
5. 加适当体积样品到96孔板的样品孔中，加标准品稀释液到20μl。
6. 各孔加入200μl BCA工作液,37℃放置20-30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，颜色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，或适当延长孵育时间。
7. 测定A562，540-595nm之间的波长也可接受。根据标准曲线计算出样品的蛋白浓度。

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。

     蛋白定量

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