Calcium Assay Kit

Produktnummer ABIN1000257

Produktdetails

Target: Calcium

Applikation: Biochemical Assay (BCA)

Proben: Serum, Urine, Saliva

Spezifität: 0.08 mg/dL (20 μM)

Produktmerkmale: 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.

Bestandteile: Reagent A: 50 mL. Reagent B: 50 mL. Calcium standard: 1 mL 20 mg/dL Ca 2+.

Benötigtes Material: 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.

Anwendungsinformationen

Applikationshinweise: 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.

Kommentare:

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

Protokoll: 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.

Aufbereitung der Reagenzien:

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

Ergebnisberechnung:

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.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Lagerung: 4 °C

Referenzen

Ou, Wu, Lai, Yang, Lo, Chiu, Bowley: "Effects of the nanostructure and nanoporosity on bioactive nanohydroxyapatite/reconstituted collagen by electrodeposition." in: Journal of biomedical materials research. Part A, Vol. 92, Issue 3, pp. 906-12, (2010) (PubMed).

Zarjou, Jeney, Arosio, Poli, Zavaczki, Balla, Balla: "Ferritin ferroxidase activity: a potent inhibitor of osteogenesis." in: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research, Vol. 25, Issue 1, pp. 164-72, (2010) (PubMed).

Ponda, Barash, Feig, Fisher, Skolnik: "Moderate kidney disease inhibits atherosclerosis regression." in: Atherosclerosis, Vol. 210, Issue 1, pp. 57-62, (2010) (PubMed).

Jung, Park, Han: "Effects of HA released calcium ion on osteoblast differentiation." in: Journal of materials science. Materials in medicine, Vol. 21, Issue 5, pp. 1649-54, (2010) (PubMed).

Chaumet-Riffaud, Martinez-Duncker, Marty, Richard, Prigent, Moati, Sarda-Mantel, Scherman, Bessodes, Mignet: "Synthesis and application of lactosylated, 99mTc chelating albumin for measurement of liver function." in: Bioconjugate chemistry, Vol. 21, Issue 4, pp. 589-96, (2010) (PubMed).

Brand, Lee, Veses, Gow: "Calcium homeostasis is required for contact-dependent helical and sinusoidal tip growth in Candida albicans hyphae." in: Molecular microbiology, Vol. 71, Issue 5, pp. 1155-64, (2009) (PubMed).

Villa-Bellosta, Sorribas: "Phosphonoformic acid prevents vascular smooth muscle cell calcification by inhibiting calcium-phosphate deposition." in: Arteriosclerosis, thrombosis, and vascular biology, Vol. 29, Issue 5, pp. 761-6, (2009) (PubMed).

Koreckij, Nguyen, Brown, Yu, Vessella, Corey: "Dasatinib inhibits the growth of prostate cancer in bone and provides additional protection from osteolysis." in: British journal of cancer, Vol. 101, Issue 2, pp. 263-8, (2009) (PubMed).

Henderson, He, Jabbari: "Concurrent differentiation of marrow stromal cells to osteogenic and vasculogenic lineages." in: Macromolecular bioscience, Vol. 8, Issue 6, pp. 499-507, (2008) (PubMed).

Warotayanont, Zhu, Snead, Zhou: "Leucine-rich amelogenin peptide induces osteogenesis in mouse embryonic stem cells." in: Biochemical and biophysical research communications, Vol. 367, Issue 1, pp. 1-6, (2008) (PubMed).

Chanda, Isayeva, Kumar, Siegal, Szafran, Zinn, Reddy, Ponnazhagan: "Systemic osteoprotegerin gene therapy restores tumor-induced bone loss in a therapeutic model of breast cancer bone metastasis." in: Molecular therapy : the journal of the American Society of Gene Therapy, Vol. 16, Issue 5, pp. 871-8, (2008) (PubMed).

He, Ma, Jabbari: "Effect of grafting RGD and BMP-2 protein-derived peptides to a hydrogel substrate on osteogenic differentiation of marrow stromal cells." in: Langmuir : the ACS journal of surfaces and colloids, Vol. 24, Issue 21, pp. 12508-16, (2008) (PubMed).

Antigendetails

Target: Calcium

Substanzklasse: Element

Hintergrund: 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.