Pierce™ BCA 蛋白检测试剂 A
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Pierce™ BCA 蛋白检测试剂 A
Thermo Scientific™

Pierce™ BCA 蛋白检测试剂 A

BCA 蛋白检测试剂 A 是 Pierce BCA 蛋白定量检测试剂盒的组成部分,后者是一种双组分、高精度,与去垢剂兼容的检测试剂,与蛋白标准品相比可用于总蛋白浓度测定。Pierce BCA了解更多信息
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货号数量
23228500 mL
232223.75 L
23221250 mL
232231 L
货号 23228
价格(CNY)
1,705.00
Each
添加至购物车
数量:
500 mL
请求批量或定制报价
价格(CNY)
1,705.00
Each
添加至购物车
BCA 蛋白检测试剂 A 是 Pierce BCA 蛋白定量检测试剂盒的组成部分,后者是一种双组分、高精度,与去垢剂兼容的检测试剂,与蛋白标准品相比可用于总蛋白浓度测定。Pierce BCA 试剂可准确测定蛋白研究中遇到的大多数样品类型的蛋白浓度。Pierce BCA 测定可用于评估全细胞裂解物、亲和柱分离组分、纯化蛋白样品的产率,以及监测工业应用中的蛋白污染。与大多数染料结合法相比,BCA 蛋白定量法受蛋白质氨基酸组成差异的影响更小,因此有更高的蛋白间定量一致性。

比较所有可用的 BCA 蛋白检测 ›

Pierce BCA 蛋白检测试剂盒(试剂 A 和 B)的特点包括:
比色法测量—可以目测评估,也可使用标准分光光度计或酶标仪 (562 nm) 进行测量
高度一致性—与染料结合法 (Bradford) 相比,蛋白间差异更小
兼容性强—在大多数离子型和非离子型去污剂的常见浓度下,性能不受影响
定量时间— 30 分钟孵育;与传统 Lowry 法相比,操作更简单,且检测速度快 4 倍
检测范围—BSA 的线性工作范围为 20 至 2000 µg/mL
高灵敏度—使用增强方案,可检测浓度低至 5 µg/mL 的样品

测定工作原理
BCA 蛋白检测法是在碱性环境下,通过蛋白将 Cu2+ 还原为 Cu1+,再通过二喹啉甲酸 (BCA) 来检测亚铜阳离子 (Cu1+)。第一步是铜与碱性环境中的蛋白质螯合,形成浅蓝色复合物,这一过程被称为双缩脲反应。在该反应(称为双缩脲反应)中,含有三个或更多氨基酸残基的肽在含酒石酸钠钾的碱性环境中与铜离子形成有色螯合复合物。

在显色反应的第二步中,BCA 与第一步形成的还原型(亚铜)阳离子反应。两分子 BCA 与一个亚铜离子螯合,产生深紫色反应产物。BCA/铜复合物是水溶性的,在 562 nm 处具有强线性吸光度,吸光度随蛋白浓度增加而升高。该复合物的灵敏度(检测下限)大约是第一个反应中淡蓝色的 100 倍。

反应很大程度上受到蛋白氨基酸序列中四个氨基酸残基(半胱氨酸、胱氨酸、酪氨酸和色氨酸)的影响。然而,与 Coomassie 染料结合法不同,通用肽骨架也会影响颜色形成,有助于尽可能减少蛋白组成差异造成的差异性。

相关产品
Pierce BCA 蛋白定量分析试剂盒
Pierce BCA 蛋白检测试剂 B
Pierce BCA 固体
仅供科研使用。不可用于诊断程序。
规格
检测BCA 蛋白定量:
描述Pierce BCA 蛋白检测试剂 A
适用于(应用)基于溶液的检测、吸光度
适用于(设备)分光光度计、酶标仪
产品线Pierce
产品类型蛋白定量检测
数量500 mL
特异性无靶标特异性
检测方法比色法
Unit SizeEach
内容与储存
在室温下储存。

常见问题解答 (FAQ)

Which Thermo Scientific protein assay is the best or the most reliable?

The choice of protein assay is dependent on preferences related to assay speed, accuracy and sensitivity, as well as interfering substances in the sample to be assayed. BCA has less protein-to-protein variation, is compatible with most detergents, and has larger working range. Pierce Bradford Plus Protein Assay Kit (Cat. Nos. 23236, A55866) is compatible with reducing sugars, is more sensitive and is faster and easier to use. For a comparison of different protein assays and compatible reagents, see our Tech Tip: Protein Quantitation Assay Compatibility Table (https://assets.thermofisher.com/TFS-Assets/LSG/Application-Notes/TR0068-Protein-assay-compatibility.pdf).

Find additional tips, troubleshooting help, and resources within our Protein Assays and Quantitation Support Center.

What substances interfere with the BCA Protein Assay?

Reducing agents, copper chelators and solutions with very high buffering capacities will interfere with the BCA assay. Reducing agents reduce the copper and will produce a high background. Copper chelators bind the copper and prevent it from being detected by the BCA reagent. High-capacity buffers prevent the BCA from reaching its optimal alkaline pH. For a complete list of compatible substances, please refer to the BCA Protein Assay Kit product instructions.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Quantitation Support Center.

What is the sample to working reagent ratio when using the BCA Protein Assay Kit?

For the Microplate Protocol the sample to working reagent (WR) ratio is 1:8 (25 µl sample plus 200 µl WR). If the sample amount is limited, a 1:20 ratio may be used (10 µl sample); in this case, however, the detection range of the assay will be limited to 125-2,000 µg/ml. For the Test Tube Procedure (Standard or Enhanced Protocols) the ratio is 1:20 (0.1 ml sample plus 2.0 ml WR). In the Micro BCA Assay Kit (for both Microplate and Test Tube Procedures), the ratio is 1:1 (1 part sample plus 1 part WR).

Find additional tips, troubleshooting help, and resources within our Protein Assays and Quantitation Support Center.

What is the detection range for the BCA Protein Assay?

For the standard protocol, the detection range is 20-2,000 µg/ml. For the enhanced test tube protocol, the detection range is 5-250 µg/ml. The Micro BCA Protein Assay Reagent will detect 0.5-20 µg/ml of protein in the test tube assay and 1-20 µg/ml for the microplate assay.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Quantitation Support Center.

How can interfering substances be eliminated from the protein sample when using the BCA Protein Assay Kit?

There are several ways to adjust a sample to be compatible with the BCA Protein Assay: • Remove the interfering substance by dialysis or gel filtration. • If the starting protein concentration of the sample is high, dilute the sample to the point that the substance no longer interferes. • Eliminate interference by copper-chelating agents by increasing the amount of copper in the working reagent (use 4 ml or 6 ml of Reagent B/100 ml of Reagent A instead of the 2 ml of Reagent B/100 ml of Reagent A that is called for in the instructions). • Precipitate sample proteins with cold acetone or trichloroacetic acid (TCA).

Find additional tips, troubleshooting help, and resources within our Protein Assays and Quantitation Support Center.