Calcium Assay Kit

Details for Product No. ABIN1000257
Biochemical Assay (BCA)
Sample Type Serum, Urine, Saliva
Specificity 0.08 mg/dL (20 μM)
Characteristics Sensitive and accurate. Use as little as 5 µL samples. Linear detection range 0.08 mg/dL (20µM) to 20 mg/dL (5mM) Ca 2+ in 96-well plate assay.
Simple and high-throughput. The procedure involves addition of a single working reagent and incubation for 3 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. Cuvet or 96-well plate assay.
Low interference in biological samples.
No pretreatments are needed. Assays can be directly performed on raw biological samples i.e., in the presence of lipid, protein and minerals such as magnesium, iron and zinc.
Components Reagent A: 50 mL. Reagent B: 50 mL. Calcium standard: 1 mL 20 mg/dL Ca 2+.
Material not included Pipeting devices and accessories (e.g. 5 µL). Clear bottom 96-well plates (e.g. Corning Costar) and plate reader. Cuvets and Spectrophotometer for measuring OD612nm.
Target Name (Antigen)
Background Quantitative determination of calcium ion Ca2+ by colorimetric (612nm) method.
Procedure: 3 min.

CALCIUM is measured to monitor diseases of the bone or calcium regulation disorders. Increased calcium levels in serum are reported in hyperparathyroidism, metastatic bone lesions and hypervitaminosis, while decreased levels are observed in hypoparathyroidism, nephrosis, rickets, steatorrhea, nephritis and calcium-losing syndromes. Urinary calcium levels aid the clinician in understanding how the kidneys handle calcium in certain diseases of the parathyroid gland. Urinary calcium levels are also essential in the medical evaluation of kidney stones. Simple, direct and automation-ready procedures for measuring calcium concentration in biological samples are becoming popular in Research and Drug Discovery. This calcium assay kit is designed to measure calcium directly in biological samples without any pretreatment. A phenolsulphonephthalein dye in the kit forms a very stable blue colored complex specifically with free calcium. The intensity of the color, measured at 612 nm, is directly proportional to the calcium concentration in the sample. The optimized formulation minimizes any interference by substances such as magnesium, lipid, protein and bilirubin.
Application Notes Direct Assays: Ca 2+ in serum, urine, saliva etc.
Drug Discovery/Pharmacology: effects of drugs on calcium metabolism.
Food and Beverages: calcium determination.
Environment: calcium determination in water and soil.

EDTA and other Ca 2+ chelators interfere with this assay. This assay can not be applied to plasma samples obtained with EDTA.

Protocol Procedure using 96-well plate:
1. Transfer 5 µL diluted standards and samples into wells of a clear bottom 96-well plate. Store diluted standards at 4°C for future use.
2. Add 200 µL working reagent and tap lightly to mix.
3. Incubate 3 min at room temperature and read optical density at 570- 650nm (peak absorbance at 612nm).

Procedure using cuvette:
1. Set up test tubes for diluted standards and Samples. Transfer 15 µL diluted Standards and samples to appropriately labeled tubes.
2. Add 1000 µL working reagent and vortex to mix. Incubate 3 min. Transfer to cuvet and read optical density at 612nm.
Reagent Preparation

Prepare enough working reagent by combining equal volumes of Reagent A and Reagent B. Equilibrate to room temperature before use.

Calculation of Results

Subtract blank OD (water, #8) from the standard OD values and plot the OD against Ca 2+ standard concentrations. Determine the slope using linear regression fitting.
Conversions: 1 mg/dL Ca 2+ equals 250 µM, 0.001% or 10 ppm.

Restrictions For Research Use only
Storage 4 °C
Supplier Images
Image no. 1 for Calcium Assay Kit (ABIN1000257) Calcium Assay Kit
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