铜含量检测试剂盒(Cuprizone, 比色法)

货号: C1507287
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级别和纯度: BioReagent ? 生物试剂级(BioReagent)—— 经测试适用于生命科学和分子生物学。适用于需要生物相容性的细胞培养、检测和生化工作。
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50T
C1507287-50T
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¥619.90
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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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2-8°C储存,避光,室温。低温运输 。请查阅批次 COA 获取详细规格。

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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 0 篇同行评审文献引用。

概述

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,通过分光光度计测定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。


C1507287
Component
50TStorage
C1507287A
铜标准 (100μg/mL)1 mL2-8℃
C1507287B
铜标准稀释液10 mLRT
C1507287C
Cu酸化液20 mLRT
C1507287D
Lea蛋白沉淀液30 mLRT. Store in the dark.
C1507287E
Cu Assay Buffer75 mL2-8℃
C1507287F
Cuprizone显色液5.5 mL2-8℃. Store in the dark.
C1507287G
ddH₂O
10 mLRT

自备仪器和试剂

离心管或试管、离心机、分光光度计、比色杯

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。

3、Cu加样
最好选用经稀盐酸处理及去离子水清洁的干燥试管或者一次性无菌聚乙烯离心管,如果无法达到上述要求,应尽量避免使用铜污染的器皿,按下表操作。
加入物 (ml)
空白管
标准管
测定管
ddH₂O
0.5—
—
铜标准 (4μg/ml)
—
0.5
—
待测样品
—
—
0.5
Cu酸化液
0.35
0.35
0.35

充分混匀,室温放置10min。

Lea蛋白沉淀液
0.50.5
0.5

充分混匀,室温放置10min,3000g离心10min,吸取各管上清液。

上述上清液
1
1
1
Cu Assay Buffer
1.4
1.4
1.4
Cuprizone显色液
0.10.1
0.1

4、Cu测定

混匀,室温静置20min,以空白调零,分光光度计620nm处检测标准管和测定管的吸光度(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 spectrophotometer. The intensity of the blue color is linearly related to the copper concentration in serum or plasma. By comparing the sample absorbance to a standard, the copper content is calculated using a formula. This assay kit demonstrates good linearity up to 62.8 μmol/L, high specificity, and considerable sensitivity. This kit is intended for research use only and is not suitable for clinical diagnosis or other purposes.

C1507287
Component
50TStorage
C1507287A
Copper Standard (100 μg/mL)
1 mL2-8℃
C1507287B
Copper Standard Diluent
10 mLRT
C1507287C
Cu Acidification Solution
20 mLRT
C1507287D
Lea Protein Precipitation Solution
30 mLRT. Store in the dark.
C1507287E
Cu Assay Buffer75 mL2-8℃
C1507287F
Cuprizone Chromogenic Solution
5.5 mL2-8℃. Store in the dark.
C1507287G
ddH₂O
10 mLRT

Materials Required but Not Provided

Centrifuge tubes or test tubes, Centrifuge, Spectrophotometer, Cuvettes

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 preferable to use dry test tubes cleaned with dilute hydrochloric acid and deionized water, or disposable sterile polyethylene centrifuge tubes. If these conditions cannot be met, strictly avoid using utensils contaminated with copper. Proceed according to the table below.

Reagent (mL)
Blank Tube
Standard Tube
Test Tube
ddH₂O
0.5—
—
Copper Standard (4 μg/mL)
—
0.5
—
Test Sample
—
—
0.5
Cu Acidification Solution
0.35
0.35
0.35
Mix thoroughly and incubate at room temperature for 10 min.
Lea Protein Precipitation Solution
0.50.5
0.5

Mix thoroughly, incubate at room temperature for 10 min, then centrifuge at 3000 × g for 10 min. Collect the supernatant from each tube.

Resulting Supernatant
1
1
1
Cu Assay Buffer
1.4
1.4
1.4
Cuprizone Chromogenic Solution
0.10.1
0.1

4. Cu Measurement

Mix well and incubate at room temperature for 20 minutes. Using the Blank tube to zero the instrument, measure the absorbance of the Standard and Test tubes at 620 nm using a spectrophotometer (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

  1. If the sample concentration is too high, dilute with deionized water and re-assay, multiplying the result by the dilution factor.

  2. Use high-purity deionized water throughout the procedure, not ordinary distilled water.

  3. Glassware should ideally be soaked in 10% hydrochloric acid for 24 hours and rinsed thoroughly with deionized water before use.

  4. Avoid copper contamination from instruments, test tubes, syringes, etc., during the experiment.

  5. The measurement wavelength can be selected between 600–640 nm with minimal impact on results.

  6. The color developed by this kit is stable and can be maintained for 1 hour at 4–20°C.

  7. Color may fade or disappear over time; complete measurements within 1 hour.

  8. Store Cu Assay Buffer sealed tightly immediately after use to prevent evaporation of active components.

  9. 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.

  10. 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.

规格

别名
络合法铜含量检测试剂盒 | 络合法铜含量检测试剂盒 | 铜检测试剂盒 | 铜比色法测试盒(络合法)
英文别名
Copper (Cu2+) Colorimetric Assay Kit (Complexing Method) | Copper (Cu2+) Colorimetric Assay Kit (Complexing Method)
规格或纯度
BioReagent
英文名称
Copper Content Assay Kit (Cuprizone, Colorimetric Method)
应用
细胞代谢
敏感性
Light-sensitive
储存与运输
储存条件
2-8°C储存,避光,室温
运输条件
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。
组分与储存
C1507287
Component
50TStorage
C1507287A
铜标准 (100μg/mL)1 mL2-8℃
C1507287B
铜标准稀释液10 mLRT
C1507287C
Cu酸化液20 mLRT
C1507287D
Lea蛋白沉淀液30 mLRT. Store in the dark.
C1507287E
Cu Assay Buffer75 mL2-8℃
C1507287F
Cuprizone显色液5.5 mL2-8℃. Store in the dark.
C1507287G
ddH₂O
10 mLRT

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ZJ26F0131493 分析证书 C1507287
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溶液计算器

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