计算溶液所需的质量、体积或浓度。
Bradford蛋白浓度测定试剂盒
有货
为什么选择此级别
BioReagent,用于蛋白分析 级 ，适用于对基线干扰要求严格的色谱和分析工作流程。
储存与运输
2-8°C储存,-20°C储存。超低温运输 。请查阅批次 COA 获取详细规格。
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SDS、COA、产品数据表及规格说明书均可下载。可通过批号查询获取批次 COA。
文献证明
在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
概述
阿拉丁生产的Bradford蛋白浓度测定试剂盒 (Bradford Protein Assay Kit) 是根据最常用的两种蛋白浓度检测方法之一Bradford法研制而成，实现了蛋白浓度测定的快速、稳定和高灵敏度，并且检测数据有很好的线性关系。
Bradford法的检测原理是考马斯亮蓝 (Coommassie Brilliant Blue) G-250与蛋白质的碱性和芳香族氨基酸特别是精氨酸 (Arginine) 在酸性介质中结合后，溶液转变为蓝色，溶液最大吸收峰从465nm迁移到595nm，颜色的变化与蛋白质浓度成正比。因此，可通过检测595nm处的吸光度对溶液中蛋白质浓度进行测定。
组分表
| B774074 | Component | 1000T | Storage |
| B774074A | Bradford蛋白浓度测定染液 | 100 mL×2 | 2-8℃ |
| B774074B | 蛋白标准 (5 mg/ml BSA) | 1 mL | -20℃ |
本产品有以下优势：
1、检测速度极快，10-80个样品只需不足10分钟即可完成。
2、灵敏度高，检测浓度下限达到25ug/ml，最小检测蛋白量达到0.5ug，待测样品体积为1-20ul。
3、在50-1000ug/ml浓度范围内有较好的线性关系。
4、Bradford法测定蛋白浓度不受绝大部分样品中的化学物质的影响。但受略高浓度的去垢剂影响。需确保SDS低于0.01%，Triton X-100低于0.05%，Tween 20, 60, 80低于0.015%。含去垢剂的样品推荐使用我们生产的BCA 蛋白定量检测试剂盒（即用型）（货号B736762）或Bradford法蛋白定量试剂盒（去垢剂兼容型）（货号B774078）。
5、 每个试剂盒可以检测1000个样品。
使用说明：
一、标准品的制备
标准试管法和标准微孔板法标准品的稀释（检测范围=100-1500 μg/ml）
| 管号 | 稀释液体积 | BSA标准品体积 | 终浓度 |
| A | 600 μl | 400 μl (从5 mg/ml管取) | 2000 μg/ml |
| B | 125 μl | 375 μl (从A管取) | 1500 μg/ml |
| C | 250 μl | 250 μl (从A管取) | 1000 μg/ml |
| D | 175 μl | 175 μl (从B管取) | 750 μg/ml |
| E | 325 μl | 325 μl (从C管取) | 500 μg/ml |
| F | 325 μl | 325 μl (从E管取) | 250 μg/ml |
| G | 325 μl | 325 μl (从F管取) | 125 μg/ml |
| H | 400 μl | 100 μl (从G管取) | 25 μg/ml |
| I | 400 μl | 0 | 0 μg/ml (空白孔) |
微量试管法和微量微孔板法标准品的稀释（检测范围=1-25 μg/ml）
| 管号 | 稀释液体积 | BSA标准品体积 | 终浓度 |
| A | 3980 μl | 20 μl (从5mg/ml管取) | 25 μg/ml |
| B | 800 μl | 3200 μl (从A管取) | 20 μg/ml |
| C | 1000 μl | 3000 μl (从B管取) | 15 μg/ml |
| D | 1000 μl | 2000 μl (从C管取) | 10 μg/ml |
| E | 1500 μl | 1500 μl (从D管取) | 5 μg/ml |
| F | 1500 μl | 1500 μl (从E管取) | 2.5 μg/ml |
| G | 4000 μl | 0 | 0 μg/ml (空白孔) |
二、蛋白浓度测定
试管测定法
A. 标准试管测定法（检测范围=100-1500 μg/ml）
1. 各取0.05ml的标准品和未知样品至相应标记的测试管中。
2. 滴加1.5ml Bradford蛋白浓度测定染液至每管中并充分混匀。
3. 可选参考: 对于大多数情况，在室温下孵育样品10分钟。
4. 设置分光光度计值于595nm并将仪器调零。然后检测所有样品的吸收值。
5. 将所有标准品及未知样品的数值中减去595nm下零孔所对应的平均值。
6. 绘制标准曲线：通过零孔平均值校正每个BSA标准品在其相应浓度下所对应的数值。再用此标准曲线判断得出每个未知样品的蛋白浓度。
B. 微量试管测定法（检测范围=1-25 μg/ml）
1. 各取1.0ml的标准品和未知样品至相应标记的测试管中。
2. 滴加1.0ml Bradford蛋白浓度测定染液至每管中并充分混匀。
3. 可选参考: 对于大多数情况，在室温下孵育样品10分钟。
4. 设置分光光度计值于595nm并将仪器调零。然后检测所有样品的吸收值。
5. 将所有标准品及未知样品的数值中减去595nm下零孔所对应的平均值。
6. 绘制标准曲线：通过零孔平均值校正每个BSA标准品在其相应浓度下所对应的数值。再用此标准曲线判断得出每个未知样品的蛋白浓度。
微孔板测定法
A. 标准微孔板测定法（检测范围=100-1500 μg/ml）
1. 各取10μl的标准品和未知样品至相应标记的微孔板中。
2. 滴加300 μl Bradford蛋白浓度测定染液至每孔混匀并充分震荡30秒。
3. 停止震荡。对于大多数情况，在室温下孵育样品10分钟。
4. 在读数仪器中测量595nm时的吸光值。
5. 将所有标准品及未知样品的数值中减去595nm下零孔所对应的平均值。
6. 绘制标准曲线：通过零孔平均值校正每个BSA标准品在其相应浓度下所对应的数值。再用此标准曲线判断得出每个未知样品的蛋白浓度。
B. 微量微孔板测定法（检测范围=1-25 μg/ml）
1. 各取150μl的标准品和未知样品至相应标记的微孔板中。
2. 滴加150 μl Bradford蛋白浓度测定染液至每孔混匀并充分震荡30秒。
3. 停止震荡。对于大多数情况，在室温下孵育样品10分钟。
4. 在读数仪器中测量595nm时的吸光值。
5. 将所有标准品及未知样品的数值中减去595nm下零孔所对应的平均值。
6. 绘制标准曲线：通过零孔平均值校正每个BSA标准品在其相应浓度下所对应的数值。再用此标准曲线判断得出每个未知样品的蛋白浓度。
注意事项：
1、将G250染液恢复到室温再使用，有利于提高检测的灵敏度，G250染液使用前请颠倒3-5次，混匀。
2、蛋白标准请在全部溶解后先混匀，再稀释成一系列不同浓度的蛋白标准。
3、为了测试准确，请在试剂加入后的5-20min内测定光吸收，因为在这段时间内颜色是最稳定的。
4、有些阳离子，如K⁺、Na⁺、Mg²⁺、(NH₄)₂SO₄、乙醇等物质不干扰测定，但大量的去污剂如TritonX-100、SDS等严重干扰测定。
5、为了您的安全和健康，请穿实验服并戴一次性手套操作。
Manufactured by Aladdin, the Bradford Protein Assay Kit is developed based on the Bradford method, one of the two most widely used protein quantification approaches. It enables rapid, stable, and highly sensitive protein concentration measurement, with excellent linearity of detection data.
Assay Principle of the Bradford Method, In an acidic medium, Coomassie Brilliant Blue G-250 binds to basic and aromatic amino acids in proteins, especially arginine. This binding turns the solution blue and shifts the maximum absorbance peak of the dye from 465 nm to 595 nm. The degree of color change is proportional to protein concentration. Therefore, the protein concentration in a sample solution can be quantified by measuring absorbance at 595 nm.
| B774074 | Component | 1000T | Storage |
| B774074A | Bradford Protein Assay Dye Reagent | 100 mL×2 | 2-8℃ |
| B774074B | Protein Standard (5 mg/ml BSA) | 1 mL | -20℃ |
Product Advantages:
1. Ultra-fast detection: Assays for 10–80 samples can be completed in less than 10 minutes.
2. High sensitivity: The lower limit of detectable concentration is 25 μg/mL, with a minimum detectable protein mass of 0.5 μg; the recommended sample loading volume ranges from 1 to 20 μL.
3. Excellent linearity within the concentration range of 50–1000 μg/mL.
4. The Bradford protein quantification assay is resistant to interference from most chemical substances in samples, but sensitive to elevated detergent concentrations. Ensure sample detergent levels comply with the limits below: SDS < 0.01%, Triton X-100 < 0.05%, Tween 20 / 60 / 80 < 0.015%. For samples containing detergents, we recommend our Ready-to-Use BCA Protein Quantitation Kit (Cat. No. B736762) or Detergent-Compatible Bradford Protein Quantitation Kit (Cat. No. B774078).
5. Each kit supports up to 1000 independent assays.
Instructions for Use:
I. Preparation of Protein Standards
Dilution Scheme for Standard Test Tube & Standard Microplate Assays (Detection range: 100–1500 μg/mL)
| Tube No. | Volume of Diluent | Volume of BSA Standard | Final Concentration |
| A | 600 μl | 400 μl (Taken from the 5 mg/ml tube) | 2000 μg/ml |
| B | 125 μl | 375 μl (Taken from tube A) | 1500 μg/ml |
| C | 250 μl | 250 μl (Taken from tube A) | 1000 μg/ml |
| D | 175 μl | 175 μl (Taken from tube B) | 750 μg/ml |
| E | 325 μl | 325 μl (Taken from tube C) | 500 μg/ml |
| F | 325 μl | 325 μl (Taken from tube E) | 250 μg/ml |
| G | 325 μl | 325 μl (Taken from tube F) | 125 μg/ml |
| H | 400 μl | 100 μl (Taken from tube G) | 25 μg/ml |
| I | 400 μl | 0 | 0 μg/ml (Blank well) |
Dilution Scheme for Micro Test Tube & Microplate Assays (Detection range: 1–25 μg/mL)
|
II. Protein Concentration Determination
Test Tube Assay
A. Standard Test Tube Assay (Detection Range: 100–1500 μg/mL)
1. Transfer 0.05 mL of each standard and unknown sample into correspondingly labeled test tubes.
2. Add 1.5 mL of Bradford protein assay dye reagent to each tube and mix thoroughly.
3. Optional guideline: Incubate samples at room temperature for 10 minutes for most applications.
4. Set the spectrophotometer wavelength to 595 nm and zero the instrument, then measure the absorbance of all samples.
5. Subtract the average absorbance value of the blank control at 595 nm from the readings of all standards and unknown samples.
6. Generate a standard curve: Correct the absorbance reading of each BSA standard at its corresponding concentration using the average blank value. Calculate the protein concentration of each unknown sample based on this standard curve.
B. Micro Test Tube Assay (Detection Range: 1–25 μg/mL)
1. Transfer 1.0 mL of each standard and unknown sample into correspondingly labeled test tubes.
2. Add 1.0 mL of Bradford protein assay dye reagent to each tube and mix thoroughly.
3. Optional guideline: Incubate samples at room temperature for 10 minutes for most applications.
4. Set the spectrophotometer wavelength to 595 nm and zero the instrument, then measure the absorbance of all samples.
5. Subtract the average absorbance value of the blank control at 595 nm from the readings of all standards and unknown samples.
6. Generate a standard curve: Correct the absorbance reading of each BSA standard at its corresponding concentration using the average blank value. Calculate the protein concentration of each unknown sample based on this standard curve.
Microplate Assay
A. Standard Microplate Assay (Detection Range: 100–1500 μg/mL)
1. Pipette 10 μL of each standard and unknown sample into correspondingly labeled wells of the microplate.
2. Add 300 μL Bradford protein assay dye reagent to each well, mix and shake vigorously for 30 seconds.
3. Stop shaking. For most samples, incubate the plate at room temperature for 10 minutes.
4. Measure the absorbance at 595 nm using a microplate reader.
5. Subtract the average absorbance value of blank wells at 595 nm from the readings of all standards and unknown samples.
6. Generate a standard curve: Correct the absorbance value of each BSA standard at its corresponding concentration with the average blank reading. Calculate the protein concentration of each unknown sample from the standard curve.
B. Micro Microplate Assay (Detection Range: 1–25 μg/mL)
1. Pipette 150 μL of each standard and unknown sample into correspondingly labeled wells of the microplate.
2. Add 150 μL Bradford protein assay dye reagent to each well, mix and shake vigorously for 30 seconds.
3. Stop shaking. For most samples, incubate the plate at room temperature for 10 minutes.
4. Measure the absorbance at 595 nm using a microplate reader.
5. Subtract the average absorbance value of blank wells at 595 nm from the readings of all standards and unknown samples.
6. Generate a standard curve: Correct the absorbance value of each BSA standard at its corresponding concentration with the average blank reading. Calculate the protein concentration of each unknown sample from the standard curve.
Precautions:
1. Equilibrate the Coomassie G-250 dye reagent to room temperature before use to improve detection sensitivity. Invert the bottle 3–5 times and mix thoroughly prior to each use.
2. Fully dissolve the BSA protein standard stock solution and mix well before serially diluting it to prepare standards of different concentrations.
3. For accurate measurements, record absorbance values within 5–20 minutes after reagent addition, as the color remains most stable during this window.
4. Most cations including K⁺, Na⁺, Mg²⁺, ammonium sulfate ((NH₄)₂SO₄), and ethanol do not interfere with the assay. However, high concentrations of detergents such as Triton X-100 and SDS will severely disrupt detection results.
5. For personal safety and health, wear a lab coat and disposable gloves during all operations.
规格
别名
Bradford法蛋白质定量检测试剂盒
规格或纯度
BioReagent, 用于蛋白分析
英文名称
Bradford Protein Assay Kit
储存与运输
储存条件
2-8°C储存,-20°C储存
运输条件
超低温运输
稳定性与储存
各组分在相应的储存条件下保质期1年。
技术文档
高级数据
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输入批号以搜索分析图谱:
通过匹配包装上的批号来查找并下载产品的 COA，每批产品都进行了严格的验证，您可放心使用！
| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 J2617016 |
分析证书 | B774074 |
| J2612494 |
分析证书 | B774074 |
| J2610171 |
分析证书 | B774074 |
| J2610166 |
分析证书 | B774074 |
| I2630010 |
分析证书 | B774074 |
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分析证书 | B774074 |
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分析证书 | B774074 |
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分析证书 | B774074 |
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分析证书 | B774074 |
| I2612048 |
分析证书 | B774074 |
| I2611511 |
分析证书 | B774074 |
| I2608597 |
分析证书 | B774074 |
| H2631424 |
分析证书 | B774074 |
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分析证书 | B774074 |
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分析证书 | B774074 |
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分析证书 | B774074 |
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分析证书 | B774074 |
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分析证书 | B774074 |
| G2630349 |
分析证书 | B774074 |
| G2625002 |
分析证书 | B774074 |
| ZJ26F0737402 |
分析证书 | B774074 |
| ZJ26F0737401 |
分析证书 | B774074 |
| G2615283 |
分析证书 | B774074 |
| G2607285 |
分析证书 | B774074 |
| G2606318 |
分析证书 | B774074 |
| G2601311 |
分析证书 | B774074 |
| F2627010 |
分析证书 | B774074 |
| F2626289 |
分析证书 | B774074 |
| F2617583 |
分析证书 | B774074 |
| F2617581 |
分析证书 | B774074 |
| ZJ25F0927136 |
分析证书 | B774074 |
| F2606002 |
分析证书 | B774074 |
| ZJ26F0535820 |
分析证书 | B774074 |
| E2623008 |
分析证书 | B774074 |
| E2623007 |
分析证书 | B774074 |
| E2614212 |
分析证书 | B774074 |
| E2607144 |
分析证书 | B774074 |
| D2623115 |
分析证书 | B774074 |
| D2620490 |
分析证书 | B774074 |
| D2611004 |
分析证书 | B774074 |
| D2611003 |
分析证书 | B774074 |
| D2610580 |
分析证书 | B774074 |
| D2610007 |
分析证书 | B774074 |
| D2601001 |
分析证书 | B774074 |
| D2601003 |
分析证书 | B774074 |
| C2630216 |
分析证书 | B774074 |
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分析证书 | B774074 |
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分析证书 | B774074 |
| C2610210 |
分析证书 | B774074 |
| A2605418 |
分析证书 | B774074 |
| L2522115 |
分析证书 | B774074 |
| L2519104 |
分析证书 | B774074 |
| L2501016 |
分析证书 | B774074 |
| K2525208 |
分析证书 | B774074 |
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分析证书 | B774074 |
| J2524747 |
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