Zinc Assay Kit

Catalog No. ABIN1000272

Product Details

Target: Zinc (Zn)

Application: Biochemical Assay (BCA)

Sample Type: Serum, Plasma (no EDTA), Urine, Saliva, Waste Water, Soil

Characteristics: Sensitive and accurate. Uses 50 µL samples. Linear detection range 0.12 µM (0.78 µg/dL) to 10 µM (65 µg/dL) zinc in 96-well assay format.
Simple and high-throughput. The procedure involves addition of a single working reagent and incubation for 30 min. Can be readily automated as a high-throughput assay for thousands of samples per day.
Improved reagent stability and versatility. The optimized formulation has greatly enhanced reagent and signal stability. Cuvette or 96-well plate assay formats possible.
Low interference in biological samples.
No pretreatments are needed.

Components: Reagent A: 50 mL. Reagent B: 1 mL. Reagent C: 1 mL. EDTA: 1 mL 100 mM. Zinc standard: 1 mL 50 µM.

Material not included: Pipeting devices and accessories. Clear bottom 96-well plates (e.g. Corning Costar) and plate reader, or cuvettes and spectrophotometer for measuring OD 425nm.

Application Details

Application Notes: Direct Assays: zinc in serum, plasma (no EDTA), urine, saliva etc.
Drug Discovery/Pharmacology: effects of drugs on zinc metabolism.
Environment: zinc determination in waste water, soil etc.

Comment:

Because the shift in the peak wavelength (from 413 nm to 425 nm) is very small, the color change is not visually evident. Physiological concentrations of other metal ions do not interfere. Zn 2+ chelators (e.g. EDTA, EGTA) should be avoided during sample preparation.

Protocol: Procedure using 96-well plate:
1. Prepare standards in deionized water. Transfer 50 µL of the Zn 2+ standards into wells of a clear flat-bottom 96-well plate. Transfer 50 µL Sample and Sample Blank (50 µL sample + 2 µL EDTA) into wells of a Add 200 µL working reagent and tap plate lightly to mix.
2. Incubate 30 min at room temperature and read optical density at 425 nm (range 420 - 426 nm).

Procedure using cuvette: Transfer 200 µL standards, sample and sample blank (200 µL Sample + 8 µL EDTA) to appropriately labeled tubes. Add 800 µL working reagent and tap lightly to mix. Incubate 30 min and read optical density at 425 nm.

Reagent Preparation:

Equilibrate all reagents to room temperature. Vortex Reagents B and C before assay. Prepare enough Working Reagent: for each assay well, mix 200 µL Reagent A, 4 µL Reagent B and 4 µL reagent C.

Sample Preparation:

Serum and plasma samples should be clear and free of turbidity or precipitates. If present, precipitates should be removed by filtration or centrifugation on a table centrifuge. Prior to assay, dilute serum or plasma samples 5-fold (n = 5) in deionized water.

Calculation of Results:

Subtract blank OD (water, #5) from the standard OD values and plot the OD against Zn 2+ standard concentrations. Calculate OD for the Sample (= ODSAMPLE - ODSAMPLE BLANK). Determine the Sample Zn 2+ concentration from the standard curve by non-linear regression fitting with a single-site saturation binding function. If the Zn 2+ concentration is higher than 10 µM, dilute sample in deionzed water. Repeat the assay and multiply the results by the dilution factor.
Conversions: 1 µM zinc equals 6.5 µg/dL or 0.065 ppm (65 ppt).

Restrictions: For Research Use only

Handling

Storage: 4 °C

References

Xu, Futakuchi, Alexander, Fukamachi, Numano, Suzui, Shimizu, Omori, Kanno, Hirose, Tsuda: "Nanosized zinc oxide particles do not promote DHPN-induced lung carcinogenesis but cause reversible epithelial hyperplasia of terminal bronchioles." in: Archives of toxicology, Vol. 88, Issue 1, pp. 65-75, (2014) (PubMed).

Morgan, Ledford, Zhou, Page: "Zinc supplementation alters airway inflammation and airway hyperresponsiveness to a common allergen." in: Journal of inflammation (London, England), Vol. 8, pp. 36, (2012) (PubMed).

Leung, Gvritishvili, Liu, Tombran-Tink: "ZIP2 and ZIP4 mediate age-related zinc fluxes across the retinal pigment epithelium." in: Journal of molecular neuroscience : MN, Vol. 46, Issue 1, pp. 122-37, (2012) (PubMed).

Kelly, Mathew, Kohler, Blass, Soybel: "Hemorrhagic shock and surgical stress alter distribution of labile zinc within high- and low-molecular-weight plasma fractions." in: Shock (Augusta, Ga.), Vol. 38, Issue 3, pp. 314-9, (2012) (PubMed).

Kelly, Mathew, Kohler, Blass, Soybel: "Redistribution of labile plasma zinc during mild surgical stress in the rat." in: Translational research : the journal of laboratory and clinical medicine, Vol. 157, Issue 3, pp. 139-49, (2011) (PubMed).

Gunasekara, Hettiarachchi, Liyanage, Lekamwasam: "Effects of zinc and multimineral vitamin supplementation on glycemic and lipid control in adult diabetes." in: Diabetes, metabolic syndrome and obesity : targets and therapy, Vol. 4, pp. 53-60, (2011) (PubMed).

Mihelich, Khramtsova, Arva, Vaishnav, Johnson, Giangreco, Martens-Uzunova, Bagasra, Kajdacsy-Balla, Nonn: "miR-183-96-182 cluster is overexpressed in prostate tissue and regulates zinc homeostasis in prostate cells." in: The Journal of biological chemistry, Vol. 286, Issue 52, pp. 44503-11, (2011) (PubMed).

Padiglia, Zonza, Atzori, Chillotti, Calò, Tepper, Barbarossa: "Sensitivity to 6-n-propylthiouracil is associated with gustin (carbonic anhydrase VI) gene polymorphism, salivary zinc, and body mass index in humans." in: The American journal of clinical nutrition, Vol. 92, Issue 3, pp. 539-45, (2010) (PubMed).

Knoell, Julian, Bao, Besecker, Macre, Leikauf, DiSilvestro, Crouser: "Zinc deficiency increases organ damage and mortality in a murine model of polymicrobial sepsis." in: Critical care medicine, Vol. 37, Issue 4, pp. 1380-8, (2009) (PubMed).

Target Details

Target: Zinc (Zn)

Alternative Name: Zinc (Zn Products)

Background: Quantitative determination of zinc ion Zn2+ by colorimetric (425nm) method.
Procedure: 30 min.

Zinc is an essential trace element and plays many key roles in metabolism. It is required for the activity of more than 300 enzymes, the structure of many proteins, and control of genetic expression. Zinc status affects basic processes of cell division, growth, differentiation, development, performance and aging through its requirement for synthesis and repair of DNA, RNA and protein. The common causes of zinc deficiency are low dietary intakes and low bioavailability. Clinical signs of zinc deficiency include acrodermatitis, low immunity, diarrhea, poor healing, stunting, hypogonadism, fetal growth failure, teratology and abortion. Zinc deficiency has now been recognized to be associated with many diseases such as malabsorption syndrome, chronic liver disease, chronic renal disease, sickle cell disease, diabetes, malignancy, and other chronic illnesses. Simple, direct and automation-ready procedures for measuring zinc concentration in biological samples are highly desirable in Research and Drug Discovery. This zinc assay kit is designed to measure zinc directly in biological samples without any pretreatment. The present method utilizes a chromogen that forms a colored complex specifically with zinc. The intensity of the color, measured at 425 nm, is directly proportional to the zinc concentration in the sample.