计算溶液所需的质量、体积或浓度。
双缩脲蛋白定量试剂盒
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
室温。常规运输 。请查阅批次 COA 获取详细规格。
质量文档
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
双缩脲是一种用于分析蛋白质的方法，双缩脲反应的原理是在呈蓝色的碱性硫酸铜溶液存在的情况下，肽键与铜离子结合，生成蓝紫色的化合物，此化合物在540nm的吸光度与肽键的数量呈正比例关系，因此可计算出蛋白质的含量，双缩脲法测蛋白浓度兼容性亦很好，不受大部分样本中其他成分的影响，但易受铜离子螯合剂的影响，另外对于血清总蛋白的双缩脲分析，胆红素、脂类、血红蛋白、葡聚糖具有一定干扰作用，双缩脲法在5～160mg/ml浓度范围内有较好的线性关系。该试剂盒仅用于科研领域，不适用于临床诊断或其他用途。
| B1507239 | Component | 500T | 1000T | Storage |
| B1507239A | 双缩脲试剂 | 100 mL | 200 mL | RT |
| B1507239B | 蛋白标准 (BSA) | 40 mg | 40 mg | RT |
| B1507239C | 蛋白标准配制液 | 1.5 mL | 3 mL | RT |
自备仪器和试剂
1、待测样品（血清、血浆、组织匀浆液）
2、离心管或小试管、水浴锅或恒温箱、酶标仪或分光光度计、96孔板或比色皿
3、生理盐水（0.9% NaCl）或PBS
实验流程
1、标准品的配制
取0.25ml蛋白标准配制液或稀释液加入到40mg蛋白标准中，充分溶解后即配制成蛋白标准溶液(160mg/ml)，配制后可立即使用，溶解后的蛋白标准溶液应-20℃保存；亦可根据需要进行稀释，如稀释至40mg/ml。特别提示：待测蛋白溶解于什么样的稀释液中，蛋白标准也宜溶解于什么样的稀释液中，例如待测蛋白溶解于蔗糖溶液中，亦取蛋白标准溶解于蔗糖溶液中，常用0.9%NaCl或PBS作为总蛋白标准品的稀释液。
2、将标准品按0、1、2、4、8、12、16、20μl加到96孔板的标准品孔，加稀释液补足至20μl。
3、加适当体积待测蛋白样本到96孔板的样品孔中，如果标准品稀释液与溶解待测蛋白样本的溶液不同，应在待测蛋白样本孔中加入20μl稀释液；如果标准品稀释液与溶解待测蛋白样本的溶液相同，无需在待测蛋白样本孔中加入20μl稀释液。
4、各孔加入200μl双缩脲试剂，室温放置10～15min。
5、测定540nm波长处的吸光值，如无540nm，510～562nm之间的波长也可。
6、根据标准曲线计算出样品的蛋白浓度。
注意事项
1、蛋白标准粉末溶解于蛋白标准配制液后，即获得蛋白标准原液，该原液中含有防腐剂，不影响后续检测，该蛋白标准原液-20℃长期保存。
2、待测蛋白和蛋白标准加入双缩脲试剂后，如果发现检测效果不佳，可以室温放置1h或60℃放置15min，颜色会随着时间的延长不断加深。
3、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铜离子螯合剂。
4、蛋白浓度低于20mg/ml时颜色反应呈不明显的蓝色，当大于40mg/ml时即可呈现明显的蓝紫色反应。
5、本试剂盒的线性范围是5~160 mg/ml，样品蛋白浓度应大于1mg/ml，以20mg/ml以上为宜，如需测定低浓度蛋白样品(1mg/ml)，推荐使用阿拉丁B665595 BCA蛋白定量试剂盒或R1491648 即用型BCA蛋白定量试剂盒。
6、本产品反应生成的蓝紫色的化合物的吸光度与试剂批次、pH值、反应温度有关，因此建议，每次测定都做标准对照。
7、氨基酸和二肽不发生双缩脲反应，三肽、寡肽和多肽与铜离子的双缩脲复合物，呈粉红色-紫红色，与蛋白质的双缩脲反应结果不同。
8、如果没酶标仪，也可以使用分光光度计测定，使用分光光度计测定蛋白浓度时，每个试剂盒可以测定的样品数量会显著减少。
9、检测中发现所有孔都呈暗紫色，可能原因是样品含有还原剂，应适当透析或稀释样品。
10、试剂开封后请尽快使用，以防影响后续实验效果。
11、蛋白标准配制成溶液后应-20℃冻存。
附录
参考标准曲线范围：
用酶标仪测定蛋白质标准1、5、10、20、40、60、80、100、120、140、160mg/ml在540nm的吸光度（未调零），空白调零后40mg/ml的吸光度为0.15~0.18，据此做出其标准曲线如下：
测定结果显示，1mg/ml的吸光度比较低，与空白管接近，因此建议测定样品浓度应大于1mg/ml，以20mg/ml以上为宜，如需测定低浓度蛋白样品(1mg/ml)，请使用阿拉丁B665595 BCA蛋白定量试剂盒或R1491648 即用型BCA蛋白定量试剂盒。
注意：对于8～16mg/ml范围内的蛋白样品，要充分考虑如EDTA、2-ME、DTT等干扰因素，其检测结果波动较大，标准品亦有波动，请注意小心精细操作。
The biuret method is an analytical technique used for protein quantification. The principle of the biuret reaction is that peptide bonds complex with copper ions in the presence of an alkaline copper sulfate solution (which is blue), generating a blue-violet compound. The absorbance of this compound at 540 nm is directly proportional to the number of peptide bonds, allowing for the calculation of protein content. The biuret method for protein concentration determination demonstrates good compatibility and is largely unaffected by other components in most samples, although it is susceptible to copper ion chelators. Additionally, for biuret analysis of serum total protein, bilirubin, lipids, hemoglobin, and dextran can cause some interference. The biuret method shows good linearity within the concentration range of 5–160 mg/mL. This kit is intended for research use only and is not suitable for clinical diagnosis or other purposes.
| B1507239 | Component | 500T | 1000T | Storage |
| B1507239A | Biuret Reagent | 100 mL | 200 mL | RT |
| B1507239B | Protein Standard (BSA) | 40 mg | 40 mg | RT |
| B1507239C | Protein Standard Diluent | 1.5 mL | 3 mL | RT |
Materials Required but Not Provided
Test samples (serum, plasma, tissue homogenate)
Centrifuge tubes or small test tubes, water bath or incubator, microplate reader or spectrophotometer, 96-well plate or cuvettes
Physiological saline (0.9% NaCl) or PBS
Experimental Procedure
1. Preparation of Standard
Add 0.25 mL of Protein Standard Diluent (or an appropriate diluent) to the 40 mg Protein Standard (BSA) vial. Dissolve completely to prepare the Protein Standard Solution (160 mg/mL). This solution can be used immediately after preparation. The dissolved Protein Standard Solution should be stored at -20°C. It can also be diluted as needed, for example to 40 mg/mL.
Special Note: The protein standard should ideally be dissolved/diluted in the same solution as the protein in the test samples. For instance, if the test protein is dissolved in a sucrose solution, the protein standard should also be dissolved in sucrose solution. Commonly, 0.9% NaCl or PBS is used as the diluent for the total protein standard.
2. Add 0, 1, 2, 4, 8, 12, 16, and 20 μL of the standard solution to the standard wells of a 96-well plate, respectively. Bring the volume in each standard well to 20 μL using the diluent.
3. Add an appropriate volume of the test protein sample to the sample wells of the 96-well plate.
If the standard diluent is different from the solution used to dissolve the test protein sample, add 20 μL of the diluent to the test sample wells.
If the standard diluent is the same as the solution used to dissolve the test protein sample, it is not necessary to add 20 μL of diluent to the test sample wells.
4. Add 200 μL of Biuret Reagent to each well. Incubate at room temperature for 10–15 minutes.
5. Measure the absorbance at 540 nm. If 540 nm is unavailable, wavelengths between 510–562 nm can also be used.
6. Calculate the protein concentration of the samples based on the standard curve.
Precautions
Dissolving the Protein Standard powder in the Protein Standard Diluent yields the stock Protein Standard Solution. This stock contains preservatives which do not interfere with the assay and can be stored long-term at -20°C.
If detection performance is suboptimal after adding Biuret Reagent to the test protein and standard, letting the reaction proceed at room temperature for 1 hour or at 60°C for 15 minutes may help, as the color continues to develop over time.
If no significant increase in absorbance or color is observed with increasing standard concentration during standard curve generation, the sample might contain copper ion chelators.
At protein concentrations below 20 mg/mL, the color reaction is a faint blue. A distinct blue-violet reaction becomes apparent at concentrations greater than 40 mg/mL.
The linear range of this kit is 5–160 mg/mL. Sample protein concentration should be greater than 1 mg/mL, preferably above 20 mg/mL. For low-concentration protein samples (~1 mg/mL), the Aladdin B665595 BCA Protein Quantification Kit or R1491648 Ready-to-Use BCA Protein Quantification Kit is recommended.
The absorbance of the blue-violet compound generated by this kit depends on the reagent batch, pH, and reaction temperature. Therefore, it is recommended to run a standard control with each assay.
Amino acids and dipeptides do not yield the biuret reaction. The biuret complexes of tripeptides, oligopeptides, and polypeptides with copper ions are pink to purplish-red, differing from the protein biuret reaction result.
If a microplate reader is unavailable, a spectrophotometer can also be used. Using a spectrophotometer significantly reduces the number of samples measurable per kit.
If all wells appear dark purple, the sample might contain reducing agents. Consider appropriate dialysis or dilution of the sample.
Use reagents promptly after opening to prevent potential effects on subsequent experimental results.
The prepared Protein Standard Solution should be stored frozen at -20°C.
Appendix: Reference Standard Curve Range
The absorbance (un-zeroed) of protein standards at 1, 5, 10, 20, 40, 60, 80, 100, 120, 140, 160 mg/mL was measured at 540 nm using a microplate reader. After zeroing with the blank, the absorbance for 40 mg/mL typically ranged from 0.15 to 0.18. The resulting standard curve is shown below:
The results show that the absorbance for 1 mg/mL is relatively low and close to the blank. Therefore, it is recommended that the sample concentration for assay be greater than 1 mg/mL, preferably above 20 mg/mL. For low-concentration protein samples (~1 mg/mL), please use the Aladdin B665595 BCA Protein Quantification Kit or R1491648 Ready-to-Use BCA Protein Quantification Kit.
Note: For protein samples in the range of 8–16 mg/mL, fully consider potential interfering factors such as EDTA, 2-ME, and DTT. Detection results for these samples may fluctuate significantly, and the standard itself might also show variability. Please perform operations carefully and precisely.
规格
别名
双缩脲法蛋白含量检测试剂盒 | 双缩脲法蛋白含量检测试剂盒 | 总蛋白（TP）比色法测试盒（双缩脲法）
英文别名
Biuret Protein Colorimetric Assay Kit | Biuret Protein Colorimetric Assay Kit
规格或纯度
BioReagent,用于蛋白分析
英文名称
Bicinchoninic acid protein quantification kit
应用
Protein Quantification
储存与运输
储存条件
室温
运输条件
常规运输
稳定性与储存
Store at room temperature long term (12 months).
组分与储存
| B1507239 | Component | 500T | 1000T | Storage |
| B1507239A | 双缩脲试剂 | 100 mL | 200 mL | RT |
| B1507239B | 蛋白标准 (BSA) | 40 mg | 40 mg | RT |
| B1507239C | 蛋白标准配制液 | 1.5 mL | 3 mL | RT |
技术文档
高级数据
质检证书（CoA,COO,BSE/TSE 和分析图谱)
C of A & Other Certificates(BSE/TSE, COO):
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 |
|---|---|---|
 ZJ26F0535653 |
分析证书 | B1507239 |
| ZJ26F0131483 |
分析证书 | B1507239 |
技术文档和文章
溶液计算器
常见问题
什么是「用于蛋白分析」？
「用于蛋白分析」表示该产品经过加工和检测，适用于分子生物学和蛋白质组学。典型规格包括：低或不可检测的核酸酶/蛋白酶活性、低内毒素、受控的微生物限度，以及高批间重现性。
什么是「BioReagent」？
「BioReagent」表示该产品经过加工和检测，适用于生物和生化应用。规格可能包括内毒素限值、微生物控制、无菌性、低 DNase/RNase/蛋白酶活性，以及在相关情况下的生物活性验证。
本产品应如何储存？
请于室温条件下保存。
本产品如何运输？
本产品按常温标准条件运输，无需温控包装。
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每个订单都会随附分析证书（COA），您也可以从账户订单历史中下载。如需订购前的COA，请联系您的客户代表，或使用PDP上的"申请COA"按钮——我们将提供代表性批次的COA。
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