BioReagent 级 ，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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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。
总蛋白(Total Protein,TP)由白蛋白和球蛋白组成，检测白蛋白的方法有双缩脲法、色氨酸法、染料结合法，其中检测白蛋白的染料结合法可采用溴甲酚绿或溴甲酚紫染料结合，上述染料对白蛋白具有高度的亲和力，通常监测染料与白蛋白结合的初速率，该速率与样品中白蛋白浓度成正比。
阿拉丁白蛋白检测试剂盒(溴甲酚紫比色法)检测原理是在酸性环境下，白蛋白分子带正电荷，与带负电荷的溴甲酚紫(Bromocresol purple，BCP)结合生成绿色复合物，在603nm处有吸收波，该复合物的吸光度与白蛋白浓度成正比，与同样处理的白蛋白标准比较，求得待测样品中白蛋白浓度，可用于人血清样本中的白蛋白含量测定，该法操作简单、方法特异，既可手工操作，又可采用自动分析仪检测，对血清清蛋白的特异性比BCG法(溴甲酚绿法)要好，不易受时间和温度变化的影响。
自备材料：
1、离心管、小试管、分光光度计、比色杯
2、蒸馏水、生理盐水或PBS
操作步骤(仅供参考)：
1、样本处理：血清、血浆样本直接取5μl检测，对于组织样本，按组织质量(g)：提取试剂【生理盐水或PBS】(mL)=1：9比例，加入9倍体积的提取试剂，冰浴下匀浆后，2500r/min离心10min，取5μl上清待检。
2、配制白蛋白标准溶液 (40mg/mL)：取0.4mL提取试剂加入到0.1mL白蛋白标准储存液(200mg/mL)中，充分混匀即成，配制后可立即使用，可分装小份-20℃保存。特别提示：待测蛋白溶解于什么样的溶液中，白蛋白标准也宜用相应的溶液稀释，例如待测蛋白溶解于蔗糖，亦用蔗糖溶液稀释白蛋白标准，一般也可以用0.9%NaCl或PBS作为稀释白蛋白标准品的溶液。
3、配制BCP试剂：按0.1mL BCP贮存液加入100mL BCP空白试剂混合即可，混合后应4℃避光保存，1个月有效。
4、白蛋白加样: 按照下表设置各管，溶液应按照顺序依次加入，并注意避免产生气泡；如果样品浓度过高，可以减少样品用量或适当稀释后再进行测定，样品的检测最好能设置平行管。
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5、白蛋白测定：顺序加入BCP试剂，并立即混匀，室温放置(30±3)s，以空白管调零，比色杯光径1cm，以分光光度计603nm测定标准管、测定管的吸光度(即A标准、A测定)。
6、(备选步骤)当遇到脂血混浊，可设“样本空白管”：取0.01mL待测样品加入2mL BCP空白试剂，以BCP空白试剂调零，读取”样本空白管”的吸光度，用测定管的吸光度减去”样本空白管”的吸光度后的净吸光度，计算白蛋白浓度。
计算
白蛋白(g/L)=A测定/A标准×白蛋白标准液浓度(g/L)
| 健康成年人 | 36～46g/L |
注意事项：
1、严重脂血对测定有干扰，应加做标本空白予以校正。
2、如果没有分光光度计，也可以使用酶标仪测定，使用酶标仪测定蛋白浓度时，每个试剂盒可以测定的样品数量可能会显著增加。
3、本法线性范围在4～80g/L呈良好线性。
4、CV可达0.45%，回收率可达99.3%～102%，平均回收率达100.5%。
5、该法测定正常样本的批间变异系数为6.3%。
6、经测定白蛋白标准溶液(40mg/ml)其吸光度减去空白吸光度差值在0.15~0.4之间；如果样品浓度过低，可能导致测定管吸光度与标准管吸光度之间差异不大，其避免方法为：增加标准品和(或)待测样品的量以提高差异性，可使标准品和(或)待测样品从10μL增加至35μL之间的量。其计算公式应调整为：白蛋白(g/L)=A测定/A标准×白蛋白标准液浓度(g/L)×(10/实际加入样品微升数)。
7、为了您的安全和健康，请穿实验服并戴一次性手套操作。
8、试剂开封后请尽快使用，以防影响后续实验效果。
有效期
6个月有效；低温运输，按要求保存。
Total Protein (TP) consists of albumin and globulin. Common methods for albumin detection include the biuret method, tryptophan method, and dye-binding method. The dye-binding method for albumin can use bromocresol green (BCG) or bromocresol purple (BCP). These dyes have high affinity for albumin, and the initial rate of dye-albumin binding is typically measured; this rate is directly proportional to the albumin concentration in the sample.
The Aladdin Albumin Content Assay Kit (Bromocresol Purple, Colorimetric Method) operates on the following principle: in an acidic environment, albumin molecules carry a positive charge and bind to negatively charged bromocresol purple (BCP) to form a green complex with an absorption peak at 603 nm. The absorbance of this complex is proportional to albumin concentration. By comparing with an identically treated albumin standard, the albumin concentration in the test sample can be determined. This kit is intended for the quantitative determination of albumin in human serum samples. The method is simple to perform and highly specific, compatible with both manual operation and automated analyzers. It exhibits greater specificity for serum albumin than the BCG method and is less susceptible to variations in time and temperature.
Materials to Be Provided by the User
1. Centrifuge tubes, test tubes, spectrophotometer, cuvettes
2. Distilled water, normal saline, or PBS
Procedure (For Reference Only)
1. Sample Preparation: For serum/plasma samples, directly use 5 μL for detection. For tissue samples, add 9 volumes of extraction solution (normal saline or PBS) per 1 volume of tissue (g/mL). Homogenize in an ice bath, centrifuge at 2500 rpm for 10 min, and collect 5 μL of the supernatant for testing.
2. Preparation of Albumin Standard Solution (40 mg/mL): Add 0.4 mL of extraction solution to 0.1 mL of Albumin Standard Stock Solution (200 mg/mL) and mix thoroughly. The solution is ready for immediate use. Aliquot and store at -20°C for long-term storage. Important Note: Dilute the albumin standard with a solution matching the one used to dissolve the test protein (e.g., sucrose solution if the sample is dissolved in sucrose). 0.9% NaCl or PBS is generally acceptable for dilution.
3. Preparation of BCP Working Reagent: Mix 0.1 mL of BCP Stock Solution with 100 mL of BCP Blank Reagent. Store the mixture at 4°C, protected from light; it is stable for 1 month.
4. Albumin Assay Setup: Prepare tubes as shown below. Add solutions sequentially and avoid bubble formation. If the sample concentration is too high, reduce the sample volume or dilute the sample before testing. Run samples in duplicate whenever possible.
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5.Albumin Measurement: Add BCP Working Reagent sequentially and mix immediately. Incubate at room temperature for 30 ± 3 seconds. Zero the spectrophotometer with the Blank Tube, then measure the absorbance of the Standard Tube and Test Tube at 603 nm using a 1-cm light path cuvette (record as Astandard and Asample .
6.(Optional) Sample Blank for Lipemic Samples: For turbid/lipemic samples, prepare a "Sample Blank Tube": add 0.01 mL of test sample to 2 mL of BCP Blank Reagent. Zero with BCP Blank Reagent and read the absorbance of the Sample Blank Tube. Calculate the net absorbance by subtracting the Sample Blank absorbance from the Test Tube absorbance for albumin concentration calculation.
Calculation
Albumin Concentration (g/L) = ( Asample / Astandard ) × Albumin Standard Concentration (g/L)
Reference Range
| Healthy Adults | 36–46 g/L |
Precautions
1. Severe lipemia interferes with the assay; correct by running a sample blank.
2. A microplate reader may be used if a spectrophotometer is unavailable; the number of tests per kit may increase significantly.
3. The assay exhibits good linearity within the range of 4–80 g/L.
4. Coefficient of variation (CV): 0.45%; recovery rate: 99.3%–102% (average: 100.5%).
5. Inter-assay CV for normal samples: 6.3%.
6. As determined experimentally, the absorbance difference (sample absorbance minus blank absorbance) of the albumin standard solution (40 mg/mL) ranges from 0.15 to 0.4. If the sample concentration is excessively low, the absorbance difference between the assay tube and standard tube will be insignificant. To avoid this problem, increase the loading volume of the standard and/or test sample to amplify absorbance discrepancy; the addition volume of standard and/or test sample can be raised from 10 μL to 35 μL accordingly. The revised calculation formula is shown below: Albumin Concentration (g/L) = ( Asample / Astandard) × Albumin Standard Concentration (g/L) × (10/Actual Sample Volume in μL)
7. Wear a lab coat and disposable gloves for personal safety.
8. Use reagents promptly after opening to avoid compromising assay performance.
Shelf Life
6 months from manufacture. Ship under cold conditions and store as specified.
| A1521771 | 组件 | 物理外观 | 100T | 储存 |
| A1521771A | BCP贮存液 | 橘色透明液体 | 0.5ml | 2-8 ℃ 避光 |
| A1521771B | 白蛋白标准储存液(200mg/ml) | 黄色透明液体 | 0.1ml | 2-8 ℃ 避光 |
| A1521771C | BCP空白试剂 | 无色透明液体 | 3×100ml | 室温 |
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ26F0535892 |
分析证书 | A1521771 |
| H2626301 |
分析证书 | A1521771 |
| H2611255 |
分析证书 | A1521771 |
| F2612211 |
分析证书 | A1521771 |
| F2603453 |
分析证书 | A1521771 |
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