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。
铜是人体重要的微量元素，是酶的重要组成部分如铜蓝蛋白（亚铁氧化酶）、超氧化物歧化酶、细胞色素氧化酶等，总含量约为90mg，有90~95%的铜与球蛋白结合，形成铜蓝蛋白；5~10%的铜与清蛋白结合，清蛋白可作为血浆铜的载体，铜的检测方法有分光光度计、原子吸收光谱法、中子活化分析法等。
检测原理：在酸性介质中与白蛋白结合的铜从白蛋白中解离出来，经沉淀蛋白质后铜离子与Cuprizone络合，形成稳定的蓝色络合物，通过酶标仪测定620nm处吸光度，形成的蓝色与血清、血浆铜成线性关系，与标准品比较，根据公式计算出铜含量，该检测试剂盒在62.8μmol/L以下线性关系良好，特异性高，较为灵敏。该试剂盒仅用于科研领域，不适用于临床诊断或其他用途。
| C1507278 | Component | 100T | Storage |
| C1507278A | 铜标准 (100μg/mL) | 1 mL | 2-8℃ |
| C1507278B | 铜标准稀释液 | 5 mL | RT |
| C1507278C | Cu酸化液 | 15 mL | RT |
| C1507278D | Lea蛋白沉淀液 | 25 mL | RT. Store in the dark. |
| C1507278E | Cu Assay Buffer | 15 mL | 2-8℃ |
| C1507278F | Cuprizone显色液 | 1.2 mL | 2-8℃. Store in the dark. |
| C1507278G | ddH₂O | 10 mL | RT |
自备仪器和试剂
1.5ml离心管或试管、离心机、酶标仪、96孔板
实验流程
1、制备样品
血浆、血清按照常规方法制备，可以直接用于该试剂盒的测定，-20℃冻存，用于Cu的检测。
2、配制铜标准工作液
取适量的铜标准 (100μg/mL) ，按铜标准(100μg/mL)：铜标准稀释液=1:24的比例配制铜标准(4μg/mL)，即为铜标准工作液；4℃避光保存，3个月有效。
| 加入物(μl) | 空白管/孔 | 标准管/孔 | 测定管/孔 |
| ddH₂O | 200 | — | — |
| 铜标准 (4μg/ml) | — | 200 | — |
| 待测样品 | — | — | 200 |
| Cu酸化液 | 140 | 140 | 140 |
充分混匀，室温放置10min。
| Lea蛋白沉淀液 | 200 | 200 | 200 |
充分混匀，室温放置10min，3000g离心10min，吸取各管上清液，取96孔板按下列步骤操作。
| 上述上清液 | 100 | 100 | 100 |
| Cu Assay Buffer | 140 | 140 | 140 |
| Cuprizone显色液 | 10 | 10 | 10 |
4、Cu测定
混匀，室温静置20min，以空白调零，酶标仪620nm处测定标准孔和测定孔的吸光度（即为A标准和A测定）。
5、结果计算
血浆、血清铜(μmol/L)=(A测定/A标准)×62.8
参数说明：
A测定：测定管的吸光度
A标准：标准管的吸光度
铜标准(4μg/ml)：铜标准(62.8μmol/L)
μg/dl=μmol/L/0.157
参考区间：
| 成年健康男性血清铜含量 | 10.99~21.98μmol/L (70~140μg/dl) |
| 成年健康女性血清铜含量 | 12.56~23.55μmol/L (80~150μg/dl) |
注意事项
1、如果样品浓度过高，应用去离子水稀释后重测，结果乘以稀释倍数。
2、实验过程中用到的水，不可用普通的蒸馏水，尽量采用高纯度的去离子水。
3、玻璃器材尽量用10%的盐酸浸泡24h，取出后用去离子水冲洗后才可以使用。
4、试验中仪器、试管、注射器等尽量避免铜污染。
5、测定波长可选600~640nm，检测结果差异不大。
6、该试剂盒呈色稳定，显色后在4~20℃可稳定1h。
7、随着时间的延长，颜色会变浅或消失，应在1h内检测完毕。
8、Cu Assay Buffer应密闭保存，用完立即盖紧盖子，防止有效成分挥发。
9、经典标准品采用铜标准(2μg/ml)，如仪器精密干扰小，可以采用铜标准(2μg/ml)，其计算公式为：血浆、血清铜(μmol/L)=(A测定/A标准)×31.4。
10、试剂开封后请尽快使用，以防影响后续实验效果。
附录
参考标准曲线范围：空白调零，通过分光光度计测定吸光度，铜标准为2μg/ml时，OD值为0.05~0.07；铜标准为4μg/ml时，OD值为0.10~0.13；通过测定铜标准为0.1、0.5、1、2、4、8、16μg/ml时的吸光度，作出其标准曲线如下：
注意：由于检测仪器和操作手法等条件的不同，参考值范围会有波动，该值仅供参考，对于要求精确计算铜含量的，可以采用标准曲线进行多点测定；根据测定经验显示，标准品浓度在0.5μg/ml以下，50μg/ml以上，标准曲线会有偏差。
Copper is an essential trace element in the human body and a key component of enzymes such as ceruloplasmin (ferroxidase), superoxide dismutase, and cytochrome oxidase. The total body copper content is approximately 90 mg, with 90–95% bound to globulin as ceruloplasmin and 5–10% bound to albumin, which serves as a plasma copper carrier. Detection methods for copper include spectrophotometry, atomic absorption spectroscopy, and neutron activation analysis.
Detection Principle
In an acidic medium, copper bound to albumin dissociates. After protein precipitation, copper ions chelate with Cuprizone to form a stable blue complex. The absorbance is measured at 620 nm using a microplate reader. The intensity of the blue color is linear with the copper concentration in serum or plasma. The copper content is calculated by comparing the sample absorbance to a standard curve. This kit demonstrates good linearity up to 62.8 μmol/L, high specificity, and considerable sensitivity. It is intended for research use only and not suitable for clinical diagnosis or other purposes.
| C1507278 | Component | 100T | Storage |
| C1507278A | Copper Standard (100 μg/mL) | 1 mL | 2-8℃ |
| C1507278B | Copper Standard Diluent | 5 mL | RT |
| C1507278C | Cu Acidification Solution | 15 mL | RT |
| C1507278D | Lea Protein Precipitation Solution | 25 mL | RT. Store in the dark. |
| C1507278E | Cu Assay Buffer | 15 mL | 2-8℃ |
| C1507278F | Cuprizone Chromogenic Solution | 1.2 mL | 2-8℃. Store in the dark. |
| C1507278G | ddH₂O | 10 mL | RT |
Materials Required but Not Provided
1.5 mL centrifuge tubes or test tubes, Centrifuge, Microplate reader, 96-well plate
Experimental Procedure
1. Sample Preparation
Plasma or serum samples prepared by standard methods can be used directly. Store at -20°C for Cu detection.
2. Preparation of Copper Standard Working Solution
Dilute the Copper Standard (100 μg/mL) with Copper Standard Diluent at a ratio of 1:24 (Copper Standard : Diluent) to obtain a Copper Standard Working Solution (4 μg/mL). Store at 4°C protected from light; stable for 3 months.
3. Cu Assay Setup
It is best to use dry test tubes cleaned with dilute hydrochloric acid and deionized water, or disposable sterile 1.5 mL polyethylene centrifuge tubes, along with a clean 96-well plate. If these conditions cannot be met, avoid using copper-contaminated utensils. Proceed according to the table below.
| Reagent (μL) | Blank Tube/Well | Standard Tube/Well | Test Tube/Well |
| ddH₂O | 200 | — | — |
| Copper Standard (4 μg/mL) | — | 200 | — |
| Test Sample | — | — | 200 |
| Cu Acidification Solution | 140 | 140 | 140 |
Mix thoroughly and incubate at room temperature for 10 min.
| Lea Protein Precipitation Solution | 200 | 200 | 200 |
Mix thoroughly, incubate at room temperature for 10 min, then centrifuge at 3000 × g for 10 min. Transfer the supernatant from each tube to a 96-well plate as follows:
| Resulting Supernatant | 100 | 100 | 100 |
| Cu Assay Buffer | 140 | 140 | 140 |
| Cuprizone Chromogenic Solution | 10 | 10 | 10 |
4. Cu Measurement
Mix well and incubate at room temperature for 20 minutes. Using the Blank to zero the instrument, measure the absorbance of the Standard and Test wells at 620 nm (recorded as Astandard and Atest ).
5. Calculation of Results
Plasma/Serum Copper (μmol/L) = (Atest / Astandard ) × 62.8
Parameter Description:
Atest : Absorbance of the test well
Astandard : Absorbance of the standard well
Copper Standard (4 μg/mL) is equivalent to 62.8 μmol/L
Unit Conversion: μg/dL = μmol/L / 0.157
Reference Interval:
| Sample Type | Serum Copper Concentration |
| Healthy Adult Males | 10.99 ~ 21.98 μmol/L (70 ~ 140 μg/dL) |
| Healthy Adult Females | 12.56 ~ 23.55 μmol/L (80 ~ 150 μg/dL) |
Precautions
If the sample concentration is too high, dilute with deionized water and re-assay, multiplying the result by the dilution factor.
Use high-purity deionized water throughout the procedure, not ordinary distilled water.
Glassware should ideally be soaked in 10% hydrochloric acid for 24 hours and rinsed thoroughly with deionized water before use.
Avoid copper contamination from instruments, test tubes, syringes, etc., during the experiment.
The measurement wavelength can be selected between 600–640 nm with minimal impact on results.
The color developed by this kit is stable and can be maintained for 1 hour at 4–20°C.
Color may fade or disappear over time; complete measurements within 1 hour.
Store Cu Assay Buffer sealed tightly immediately after use to prevent evaporation of active components.
The classical standard uses a Copper Standard (2 μg/mL). If instrument precision is high and interference is low, this standard can be used, and the calculation formula becomes: Plasma/Serum Copper (μmol/L) = (Atest / Astandard ) × 31.4.
Use reagents promptly after opening to prevent potential effects on subsequent experiments.
Appendix
Reference Standard Curve Range: (Zeroed with Blank)
Absorbance (OD) for the 2 μg/mL Copper Standard typically ranges from 0.05 to 0.07 when measured by spectrophotometer.
Absorbance (OD) for the 4 μg/mL Copper Standard typically ranges from 0.10 to 0.13.
The standard curve obtained by measuring the absorbance of copper standards at 0.1, 0.5, 1, 2, 4, 8, and 16 μg/mL is shown below:
Note: Reference ranges may vary due to differences in detection instruments and operational techniques. These values are for reference only. For precise copper quantification, multi-point calibration using a standard curve is recommended. Empirical data suggests deviations in the standard curve may occur for standard concentrations below 0.5 μg/mL or above 50 μg/mL.
| C1507278 | Component | 100T | Storage |
| C1507278A | 铜标准 (100μg/mL) | 1 mL | 2-8℃ |
| C1507278B | 铜标准稀释液 | 5 mL | RT |
| C1507278C | Cu酸化液 | 15 mL | RT |
| C1507278D | Lea蛋白沉淀液 | 25 mL | RT. Store in the dark. |
| C1507278E | Cu Assay Buffer | 15 mL | 2-8℃ |
| C1507278F | Cuprizone显色液 | 1.2 mL | 2-8℃. Store in the dark. |
| C1507278G | ddH₂O | 10 mL | RT |
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 |
|---|---|---|
 H2611254 |
分析证书 | C1507278 |
| ZJ26F0131492 |
分析证书 | C1507278 |
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