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产品简介：
碱性条件下，蛋白将Cu2+还原为Cu+， Cu+与BCA试剂形成紫蓝色的络合物，测定其在562nm处的吸收值，并与标准曲线对比，即可计算待测蛋白的浓度。常用浓度的去垢剂SDS，Triton X-100，Tween不影响检测结果，但受螯合剂(EDTA, EGTA)、还原剂（DTT，巯基乙醇）和脂类的影响。实验中，若发现样品稀释液或裂解液本身背景值较高，可试用Bradford蛋白浓度测定试剂盒。
操作说明：
一. 微孔酶标仪法
1. 配制工作液：根据标准品和样品数量，按50体积BCA试剂加1体积Cu试剂（50:1）配制成BCA工作液，充分混匀(混合时可能会有浑浊，但混匀后就会消失)。BCA工作液室温24小时内稳定。
2. 稀释标准品：取10μL BSA标准品用PBS稀释至100μL（样品一般可用PBS稀释），使终浓度为0.5mg/mL。将标准品按0, 2, 4, 6, 8, 12, 16, 20μL加到96孔板的蛋白标准品孔中，加PBS补足至20μL。
3. 将样品作适当稀释（最好多做几个梯度，如作2倍、4倍、8倍稀释），加20μL到96孔板的样品孔中。由于移液器在取小量样品时误差偏大，标准线前面的点可能不很准确，所以尽可能的让样品点落在标准线1/2后。
4. 各孔加入200μL BCA工作液，37℃放置15-30分钟。用酶标仪测定A562nm，根据标准曲线计算出蛋白浓度。使用温箱孵育时，应注意防止因水分蒸发影响检测结果。
二. 分光光度计法 如没有酶标仪，可用分光光度计在离心管中混匀后加入比色皿中比色。 步骤如下：
1. 配制工作液: 根据标准品和样品数量，按50体积BCA试剂加1体积Cu试剂（50:1）配制成BCA工作液，充分混匀(混合时可能会有浑浊，但混匀后就会消失)。BCA工作液室温24小时内稳定。
2. 稀释标准品：取100μL BSA标准品用PBS稀释至1mL（样品一般可用PBS稀释），使终浓度为0.5mg/mL。
3. 取八支（或者更多）5mL离心管，标上号，按下表加入试剂。
4.37℃放置15-30分钟。用分光光度计测562nm处吸光值，根据标准曲线计算出蛋白浓度。
注意事项：
1. 长期不用时，Cu试剂与PBS稀释液可置于2-8℃保存，如发现细菌污染则应丢弃。BCA试剂在低温条件下出现结晶沉淀时，可37℃温育使其完全溶解, 不影响使用。
2. 样品中若含有较多干扰物质时，请采用Bradford蛋白浓度测定试剂盒。
3. 为了您的安全和健康，请穿实验服并戴一次性手套操作。
注意事项
Considerations
1.本产品仅供科研使用。请勿用于医药、临床诊断或治疗，食品及化妆品等用途。请勿存放于普通住宅区。
2.为了您的安全和健康，请穿好实验服并佩戴一次性手套和口罩操作。
3.实验结果可由多种因素影响，相关处理只限于产品本身，不涉及其他赔偿。
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相关标记
Conjugate
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论文引用
Paper Citation
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