Name: BCA蛋白定量试剂盒
SKU: B665595
Availability: InStock

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，适用于对基线干扰要求严格的色谱和分析工作流程。

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文献证明

在色谱分析、有机合成和交叉偶联反应领域已被 7 篇同行评审文献引用。

概述

BCA（Bicinchoninic acid）法是⽬前应⽤⽐较⼴泛的蛋⽩质浓度测定⽅法。基于双缩脲反应，即在碱性环境下蛋⽩质将 Cu2+还原成 Cu+，产⽣⼀种紫蓝⾊复合物，在 562nm 处有⾼的吸光值，该反应产物的量与蛋⽩质浓度成正⽐。BCA 蛋⽩浓度测定法实现了蛋⽩质浓度测定的简便、灵敏、快速和稳定性。试剂盒中提供的蛋⽩标准品为⽤户制作标准曲线提供了便利。该 BCA 蛋⽩浓度测定试剂盒可⽤于⽐⾊⽫法检测，也可⽤于微孔板法检测。前者虽需较⼤量（100μL）的蛋⽩样品，但由于其在检测中使⽤蛋⽩样品与 BCA ⼯作液的⽐率为 1:20（v/v），从⽽降低⼲扰物质带来的影响。后者操作简单⽅便，仅需少量（10-25μL）的蛋⽩样品。不过，由于其在检测中使⽤蛋⽩样品与 BCA ⼯作液的⽐率为 1:8（v/v），某种程度上限制⼲扰物质的承受浓度以及降低最低检测⽔平。

组分

组分	规格 500T	储存
BCA 试剂 A	100mL	4℃
BCA 试剂 B	3mL	4℃
蛋白标准品（BSA）	2×1mL（5mg/mL）	-20℃
PBS 溶液	10mL	4℃

产品特点

1）灵敏度高，最小检测蛋白量可达 0.2μg，检测浓度下限达到 10μg/mL(在 20~1000μg/ml 浓度范围内有较好的线性关系）。
2）速度快，比一般的 BCA 蛋白浓度测定试剂盒显色所用时间短。比传统的Lowry 法检测速度约快 4 倍。
3）线性范围广， 20-1000μg/mL 浓度范围内有较好的线性范围。
4) 不受大部分样品中的化学物质的影响, BCA 法测定蛋白浓度的最大优点是蛋白浓度的测定可以耐受高浓度的去垢剂, 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%。

5) Bradford 法与 BCA 法的对比

方法	Bradford 法	BCA 法
灵敏度	1~5 μg	0.5~20 μg
时间	5~15 min	40~60 min
波长	595 nm	562 nm
检测不同类别蛋白质	变异系数较高	变异系数较低
吸光稳定性	1 h 内稳定	随时间变化

操作说明

BCA 工作液配制：

将试剂 A 和试剂 B 按照体积比 50:1 比例混合配成 BCA 工作液。取 50mL 试剂 A 与 1mL 试剂 B 混合，配成 51mL BCA 工作液。两者混合时会有沉淀形成，彻底混匀后沉淀消失。
微孔板测定程序：（工作范围 20-2000 μg/ml）
1. 蛋白标准品配制：室温完全溶解蛋白标准品，取 20µl 5mg/ml BSA 蛋白标准溶液用 PBS 溶液稀释至 100µl 使其终浓度为 1.0 mg/ml。
2. 按照下表配制 BSA 标准测定溶液：

编号	0	1	2	3	4	5	6	7	8
/	1 mg/ml BSA 标准溶液 μL	1 mg/ml BSA 标准溶液 μL	1 mg/ml BSA 标准溶液 μL	1 mg/ml BSA 标准溶液 μL	1 mg/ml BSA 标准溶液 μL	1 mg/ml BSA 标准溶液 μL	1 mg/ml BSA 标准溶液 μL	5 mg/ml BSA 标准溶液 μL	5 mg/ml BSA 标准溶液 μL
BSA 标准溶液 µl	0	0.5	2.5	5	10	15	20	6	8
PBS 溶液 μl	20	19.5	17.5	15	10	5	0	14	12
BSA 终浓度 μg/ml	0	25	125	250	500	750	1000	1500	2000
总体积 µl	20 µl	20 µl	20 µl	20 µl	20 µl	20 µl	20 µl	20 µl	20 µl

3. 将适当体积的待测样品加入到微孔板中，并用 PBS 补足到 20 µl
4. 向微孔板中加入 200μL BCA 工作液混匀, 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, 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。
5. 测定 562 nm 处的吸光值，并记录读数；以不含 BSA 的样品的光吸收值作为空白对照。
6. 以 A562 为纵坐标, BSA 含量为横坐标, 绘制标准曲线, 计算样品中的蛋白浓度. 如果蛋白浓度不在标准曲线范围内, 请稀释样品后重新测定。
试管测定程序：（工作范围 20-1000 μg/ml）
1. 蛋白标准品配制：
室温完全溶解蛋白标准品, 取 150µL 5mg/ml BSA 蛋白标准溶液,加入 600μLPBS 溶液稀释至 750µl，使其终浓度为 1.0 mg/ml。
2、按照下表配制 BSA 标准测定溶液：

编号	0	1	2	3	4	5	6	7	8
/	1 mg/ml BSA 标准溶液 μl	1 mg/ml BSA 标准溶液 μl	1 mg/ml BSA 标准溶液 μl	1 mg/ml BSA 标准溶液 μl	1 mg/ml BSA 标准溶液 μl	1 mg/ml BSA 标准溶液 μl	1 mg/ml BSA 标准溶液 μl	5 mg/ml BSA 标准溶液 μl	5 mg/ml BSA 标准溶液 μl
BSA 标准溶液 µl	0	2.5	12.5	25	50	75	100	30	40
PBS 溶液 μl	100	97.5	87.5	75	50	25	0	70	60
BSA 终浓度 μg/ml	0	25	125	250	500	750	1000	1500	2000
总体积 µl	100 µl	100 µl	100 µl	100 µl	100 µl	100 µl	100 µl	100 µl	100 µl

3. 将适当体积的待测样品加入到试管中，并用 PBS 补足到 100 µl；
4. 向试管中加入 2ml BCA 工作液，混匀， 37 ℃放置 30 分钟；
5 、6 步骤同上

注意事项

1）BCA 蛋⽩测定法测定时颜⾊会随着时间的延⻓不断加深，显⾊反应的速度和温度有关，需注意保持定时和定温，以确保精确定量。
2）⻓期保存，若发现 BCA 试剂 A 或者试剂 B 有沉淀发⽣。请 37ºC 温育并伴随搅拌促使其充分溶解。如发现细菌污染，则应丢弃。
3）实验操作规范，提⾼上样量的精确度。
4）每次测定都需做相应的标准曲线，因为显⾊反应与温度和时间的变化有关，精准蛋⽩定量宜每次都做标准曲线。
5）本公司所有产品仅限用于研发，必须专业人员操作同时佩戴口罩/手套/实验服并遵守生物实验室安全操作规程！

The BCA (Bicinonic acid) method is a widely used method for measuring egg protein concentration. Based on the biuret reaction, which involves the reduction of Cu2+to Cu+by egg white in an alkaline environment, a purple blue complex is produced, with a high absorbance value at 562nm. The amount of the reaction product is positively correlated with the concentration of egg white. The BCA protein concentration determination method has achieved the simplicity, sensitivity, speed, and stability of protein concentration determination. The egg white standard provided in the reagent kit provides convenience for users to create standard curves. The BCA egg protein concentration determination kit can be used for nucleic acid detection or for microplate detection. The former requires a relatively large amount (100 μ L) of egg samples, but due to its use in detection, the ratio of egg samples to BCA working solution is 1:20 (v/v), thereby reducing the impact of interfering substances. The latter is easy to operate and only requires a small amount (10-25 μ L) of egg yolk sample. However, due to its 1:8 (v/v) ratio between the egg white sample and BCA working solution during detection, it to some extent limits the tolerance concentration of interfering substances and reduces the minimum detection level.

Component

Composition	Specifications 500T	Storage
BCA reagent A	100mL	4℃
BCA reagent B	3mL	4℃
Protein standards（BSA）	2×1mL（5mg/mL）	-20℃
PBS Solution	10mL	4℃

Product features
1) High sensitivity, with a minimum detection protein content of 0.2 μ g and a detection concentration lower limit of 10 μ g/mL (with a good linear relationship in the concentration range of 20~1000 μ g/mL).
2) Fast speed and shorter color development time than typical BCA protein concentration determination kits. The detection speed is about 4 times faster than the traditional Lowry method.
3) The linear range is wide, with a good linear range within the concentration range of 20-1000 μ g/mL.
4) The biggest advantage of using BCA method to determine protein concentration is that it is not affected by most chemical substances in the sample. The determination of protein concentration can tolerate high concentrations of descaling agents and is compatible with up to 5% SDS in the sample; 5% Triton X -100; 5% Tween 20, 60, 80. However, due to the influence of chelating agents and slightly higher concentrations of reducing agents, it is necessary to ensure that EDTA is below 10mM, there is no EGTA, and dithiothreitol is below 1mM; β - mercaptoethanol is less than 0.01%. Please refer to Appendix 1 of 7 for details.
5) Comparison between Bradford method and BCA method

Method	Bradford method	BCA method
Sensitivity	1~5 μg	0.5~20 μg
Time	5~15 min	40~60 min
Wavelength	595 nm	562 nm
Detecting proteins of different categories	High coefficient of variation	Low coefficient of variation
Absorbing stability	Stable within 1 hour	Over time

Operating instructions
BCA working solution preparation:
Mix reagent A and reagent B in a volume ratio of 50:1 to prepare BCA working solution. Mix 50mL of reagent A with 1mL of reagent B to prepare 51mL of BCA working solution. When the two are mixed, precipitation will form, and after thorough mixing, the precipitation will disappear.
Microporous plate measurement program: (working range 20-2000 μ g/ml)
1. Preparation of protein standard: Dissolve the protein standard completely at room temperature, take 20 µ l 5mg/ml BSA protein standard solution and dilute it with PBS solution to 100 µ l to achieve a final concentration of 1.0 mg/ml.
2. Prepare BSA standard measurement solution according to the following table:

Number	0	1	2	3	4	5	6	7	8
/	1 mg/ml BSA Standard solution μL	1 mg/ml BSA Standard solution μL	1 mg/ml BSA Standard solution μL	1 mg/ml BSA Standard solution μL	1 mg/ml BSA Standard solution μL	1 mg/ml BSA Standard solution μL	1 mg/ml BSA Standard solution μL	5 mg/ml BSA Standard solution μL	5 mg/ml BSA Standard solution μL
BSA Standard solution µl	0	0.5	2.5	5	10	15	20	6	8
PBS Solution μl	20	19.5	17.5	15	10	5	0	14	12
BSA Final concentration μg/ml	0	25	125	250	500	750	1000	1500	2000
Total volume µl	20 µl	20 µl	20 µl	20 µl	20 µl	20 µl	20 µl	20 µl	20 µl

3. Add an appropriate volume of the test sample to the microplate and supplement it with PBS to 20 µ l
4. Add 200 μ L of BCA working solution to the microplate and mix well, then let it stand at 37 ℃ for 30 minutes; Note: It can also be left at room temperature for 2 hours or at 60 ℃ for 30 minutes. When measuring protein concentration using the BCA method, the color will deepen over time, and the color reaction will accelerate with increasing temperature. If the concentration is low, it is suitable to incubate at a higher temperature or extend the incubation time appropriately.
5. Measure the absorbance value at 562 nm and record the reading; Use the light absorption value of the sample without BSA as the blank control.
6. Draw a standard curve with A562 as the vertical axis and BSA content as the horizontal axis, and calculate the protein concentration in the sample If the protein concentration is not within the standard curve range, please dilute the sample and retest.
Test tube testing procedure: (working range 20-1000 μ g/ml)
1. Preparation of protein standards:
Dissolve the protein standard completely at room temperature, take 150 µ L 5mg/ml BSA protein standard solution, add 600 μ L LPBS solution and dilute to 750 µ l to achieve a final concentration of 1.0 mg/ml.
2. Prepare BSA standard measurement solution according to the following table:

Number	0	1	2	3	4	5	6	7	8
/	1 mg/ml BSA Standard solution μl	1 mg/ml BSA Standard solution μl	1 mg/ml BSA Standard solution μl	1 mg/ml BSA Standard solution μl	1 mg/ml BSA Standard solution μl	1 mg/ml BSA Standard solution μl	1 mg/ml BSA Standard solution μl	5 mg/ml BSA Standard solution μl	5 mg/ml BSA Standard solution μl
BSA Standard solution µl	0	2.5	12.5	25	50	75	100	30	40
PBS Solution μl	100	97.5	87.5	75	50	25	0	70	60
BSA Final concentration μg/ml	0	25	125	250	500	750	1000	1500	2000
Total volume µl	100 µl	100 µl	100 µl	100 µl	100 µl	100 µl	100 µl	100 µl	100 µl

3. Add an appropriate volume of the sample to be tested into the test tube and supplement it with PBS to 100 µ l;
4. Add 2ml of BCA working solution to the test tube, mix well, and let it stand at 37 ℃ for 30 minutes;
5. 6 steps as above

Matters needing attention

1) When using the BCA egg size determination method, the color will deepen over time, and the speed of the color reaction is related to temperature. It is important to maintain timing and temperature to ensure accurate quantification.
2) If BCA reagent A or reagent B is found to have precipitates, it should be stored for a period of time. Please incubate at 37 º C and stir to fully dissolve. If bacterial contamination is found, it should be discarded.
3) Standardize experimental operations to improve the accuracy of sample loading.
4) Each measurement requires a corresponding standard curve, as the visible reaction is related to changes in temperature and time. Accurate egg quantification should be done with a standard curve every time.
5) All products of our company are limited to research and development purposes only, and must be operated by professional personnel while wearing masks/gloves/lab clothes and complying with the safety operating procedures of the biological laboratory!

规格

英文名称

BCA Protein Assay Kit

储存与运输

储存条件

2-8°C储存

运输条件

低温运输

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批号(Lot Number)	证书类型	货号
H2625856	分析证书	B665595
F2617618	分析证书	B665595
C2613002	分析证书	B665595
A2614501	分析证书	B665595
K2521084	分析证书	B665595
J2513167	分析证书	B665595
G2402024	分析证书	B665595

技术文档和文章

b665595_bca protein assay kit.pdf

b665595_bca蛋白定量试剂盒.pdf

Western Blot

目录：技术文章

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蛋白质印迹

Western Blot

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Western blot

Western Blot常见问题

目录：常见问题(FAQs)

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WB Western blot

Common Issues for Western Blot (WB)

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WB(Western Blot)

使用磁性琼脂糖微球纯化抗体

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抗体纯化磁性琼脂糖微球

Purifying Antibodies with Magnetic Agarose Beads

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Antibody Purification Magnetic agarose microspheres

常见的11种蛋白质浓度测定方法

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蛋白质浓度测定方法

11 Common Methods for Protein Concentration Determination

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Protein Concentration Determination

DNA Pull-Down实验操作规程

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DNA Pull-Down

DNA Pull-Down Standard Operating Procedure (SOP)

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DNA Pull-Down

CK、CK-MB与CK-MM检测在组织损伤判读中的意义

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同工酶组织损伤肌酸激酶 CK CK-MB CK-MM 骨骼肌损伤心肌损伤

Significance of CK, CK-MB, and CK-MM Testing in the Interpretation of Tissue Injury

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Isoenzyme CK CK-MB CK-MM creatine kinase tissue injury skeletal muscle injury myocardial injury

抗氧化酶防御体系中SOD、CAT与GSH-Px的联合检测策略

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谷胱甘肽氧化应激活性氧过氧化氢 SOD 抗氧化酶 CAT GSH-Px 联合检测

Combined Detection Strategy for SOD, CAT, and GSH-Px in the Antioxidant Enzyme Defense System

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glutathione Oxidative Stress Reactive oxygen species Hydrogen peroxide SOD Antioxidant Enzymes CAT GSH-Px combined detection

丙二醛含量检测的原理、方法选择与结果判读

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HPLC 氧化应激 SOD 脂质过氧化 CAT GSH-Px 4-HNE MDA 丙二醛 TBA法 TBARS 硫代巴比妥酸

Principles, Method Selection, and Result Interpretation for Malondialdehyde Content Detection

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HPLC Oxidative Stress SOD lipid peroxidation CAT GSH-Px 4-HNE MDA TBARS Malondialdehyde TBA method Thiobarbituric acid

溶液计算器

摩尔浓度计算器

计算溶液所需的质量、体积或浓度。

质量

浓度

体积

分子量

g/mol

稀释计算器

计算配制储备溶液所需的稀释度。

浓度1（C1）

体积 1 (V1)

浓度 2 (C2)

体积 2 (V2)

复溶计算器

复溶计算器允许您快速计算试剂的体积，以复溶小瓶。只需输入试剂的质量和目标浓度，计算器将确定其余部分。

体积（添加到小瓶中）

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所需的复溶浓度

大包装与定制包装 需要更大包装或定制规格？请联系我们的团队获取专属报价。

BCA蛋白定量试剂盒

储存与运输

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文献证明

概述

规格

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