Copper Assay Kit

Catalog No. ABIN1000260

Product Details

Target: Copper (COP)

Application: Biochemical Assay (BCA)

Specificity: 7 μg/dL (1.0 μM)

Characteristics: Sensitive and accurate. Linear detection range 7 µg/dL (1.0 µM) to 300 µg/dL (47 µM) copper in 96-well plate assay.
Simple and high-throughput. The simple procedure can be readily automated as a high-throughput assay in 96-well plates for thousands of samples per day.
Improved reagent stability and versatility. The optimized formulation has greatly enhanced reagent and signal stability. Cuvet or 96-well plate assay.

Components: Reagent A: 10 mL. Reagent B: 1.5 mL. Reagent C: 40 mL. Copper Standard: 1 mL 1.5 mg/dL Cu2+.

Material not included: Pipeting devices and accessories. For 96-well plate assays: clear flat-bottom 96-well plates and plate reader. For cuvet assays: spectrophotometer and cuvets for measuring OD at 356-362nm.

Application Details

Application Notes: Direct Assays: biological, environmental, food and beverage samples.
Drug Discovery/Pharmacology: effects of drugs on Cu metabolism.

Protocol: Note: metal chelators (e.g. EDTA) interfere with this assay and should be avoided in sample preparation.

Procedure using 96-well plate:
1. Standards: transfer 100 µL dH2O into one Eppendorf tube labeled Blank. Into another tube labeled Standard, mix 20 µL1.5 mg/dL Standard and 80 µL dH2O (final 300 µg/dL Cu 2+ ). Samples: transfer 100 µL samples into separate tubes. Add 35 µL Reagent A (trichloroacetic acid) to each tube and mix by vortexing. If samples contain protein (e.g. serum/plasma), precipitates form. Centrifuge tubes for 2 min at 14,000 rpm and use clear supernatant for assay. For samples that do not contain protein, the mixture remains clear and centrifugation is not necessary. Transfer 100 µL Blank, Standard and Sample into separate wells of a clear flat-bottom 96-well plate.
2. For each assay well, prepare Working Reagent by mixing 5 µL Reagent B and 150 µL Reagent C. Transfer 150 µL Working Reagent to each well and tap plate to mix thoroughly.
3. Incubate 5 min at room temperature and read optical density at 356- 362nm (peak absorbance at 359nm). Note: if sample OD values are higher than the OD value for the 300µg/dL Standard, dilute sample in dH2O and repeat assay. Multiply the results by the dilution factor.

Procedure using cuvette: Prepare standards and samples as for 96-well assay procedure.
1. Transfer 400 µL Standards and Samples into separate cuvets.
2. Add 600 µL Working Reagent. Mix by pipetting.
3. Incubate 5 min at room temperature and read optical density at 356-362nm (peak absorbance at 359nm).

Restrictions: For Research Use only

Handling

Storage: 4 °C

References

Piret, Vankoningsloo, Mejia, Noël, Boilan, Lambinon, Zouboulis, Masereel, Lucas, Saout, Toussaint: "Differential toxicity of copper (II) oxide nanoparticles of similar hydrodynamic diameter on human differentiated intestinal Caco-2 cell monolayers is correlated in part to copper release and shape." in: Nanotoxicology, Vol. 6, Issue 7, pp. 789-803, (2012) (PubMed).

Bartnikas: "Known and potential roles of transferrin in iron biology." in: Biometals : an international journal on the role of metal ions in biology, biochemistry, and medicine, Vol. 25, Issue 4, pp. 677-86, (2012) (PubMed).

Philips, Samuel, Parakandi, Gopal, Siomyk, Ministro, Thompson, Borkow: "Beneficial regulation of fibrillar collagens, heat shock protein-47, elastin fiber components, transforming growth factor-β1, vascular endothelial growth factor and oxidative stress effects by copper in dermal fibroblasts." in: Connective tissue research, Vol. 53, Issue 5, pp. 373-8, (2012) (PubMed).

Piret, Jacques, Audinot, Mejia, Boilan, Noël, Fransolet, Demazy, Lucas, Saout, Toussaint: "Copper(II) oxide nanoparticles penetrate into HepG2 cells, exert cytotoxicity via oxidative stress and induce pro-inflammatory response." in: Nanoscale, Vol. 4, Issue 22, pp. 7168-84, (2012) (PubMed).

Lull, Carkaci-Salli, Freeman, Myers, Midgley, Thomas, Kimatian, Vrana, Undar: "Plasma biomarkers in pediatric patients undergoing cardiopulmonary bypass." in: Pediatric research, Vol. 63, Issue 6, pp. 638-44, (2008) (PubMed).

Target Details

Target: Copper (COP)

Alternative Name: Copper

Target Type: Element

Background: Quantitative determination of copper(II) ion Cu2+ at 359nm.
Procedure: 10 min.

Copper is an essential trace element. Copper-containing enzymes play important roles in iron and catecholamine metabolism, free radical scavenging, and in the synthesis of hemoglobin, elastin and collagen. Copper is mainly present in caeruloplasmin in the liver. Low levels of copper have been associated with mental retardation, depigmentation, anaemia, hypotonia and scorbutic changes in bone. Levels of copper are key diagnostic indicator of diseases such as Wilson's disease, microcytic hypochromic anaemia and bone disease due to reduced collagen synthesis. Simple, direct and automation-ready procedures for measuring copper concentrations find wide applications in research, drug discovery and environmental monitoring. This copper assay kit is designed to measure copper with no or minimal sample treatment. The improved method utilizes a chromogen that forms a colored complex specifically with copper ions. The intensity of the color, measured at 359nm, is directly proportional to copper concentration in the sample. The optimized formulation substantially reduces interference by substances in the raw samples.