Calcium Colorimetric Assay Kit

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Antigen
Reactivity
Chemical
1
Application
Biochemical Assay (BCA)
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Purpose Colorimetric Calcium Assay Kit utilizes the chromogenic complex (λ = 575 nm) formed between calcium ions and 0-cresolphthalein to provide a simple assay in the physiologically important range of calcium concentration 0.4-100 mg/dL (0.1-25 mM).
Sample Type Cell Lysate, Plasma, Serum, Tissue Samples
Detection Method Colorimetric
Specificity Colorimetric Calcium Assay Kit utilizes the chromogenic complex (lambda = 575 nm) formed between calcium ions and 0-cresolphthalein to provide a simple assay in the physiologically important range of calcium concentration 0.4-100 mg/dL (0.1-25 mM).
Characteristics Calcium Colorimetric Assay Kit: Measure Calcium Level in a variety of Samples such as Serum, Urine etc. Convenient & Sensitive Colorimetric Assay.
Components Calcium Assay Buffer
Chromogenic Reagent
Calcium Standard (500 mM)
Target Name (Antigen) Calcium
Target Type Chemical
Background Calcium is essential for all living organisms, where Ca²ᶧ, sequestration and release into and out of the cytoplasm functions as a signal for many cellular processes. 99 % of calcium is found in bones and teeth with the remaining 1 % found in the blood and soft tissue. Serum calcium levels are tightly controlled (8.4-11.4 mg/dL) and any variation outside this range can have serious effects. Calcium plays a role in mediating the constriction and relaxation of blood vessels, nerve impulse transmission, muscle contraction, and hormone secretion. Calcium ion channels control the migration of calcium ions across cell membranes, permitting the activation and inhibition of a wide variety of enzymes. Causes of low calcium levels include chronic kidney failure, vitamin D deficiency, and low blood magnesium levels that can occur in severe alcoholism.
Application Notes The kit can detect calcium concentration 0.4-100 mg/dL (0.1-25 mM).
Comment

Further details regarding sample type: Cell and tissue lysates, culture media, urine, plasma and serum, as well as many other biological fluids

Protocol 1. Standard Curve Preparations: Dilute the Calcium Standard to 5 mM (20 mg/dL) by adding 10 µL of the 500 mM Standard to 990 µL of dH2O, mix well. Add 0, 2, 4, 6, 8, 10 µL into a series of wells to give 0, 0.4, 0.8, 1.2, 1.6, 2.0 µg calcium per well. Bring the volume to total 50 µL with dH2O.
2. Sample Preparation: Serum or urine samples can be used directly in this assay. Place 10 µL samples in wells in a 96-well plate. For other liquid samples, add 2 - 50 µL sample into individual well. Bring the total volume to total 50 µL with dH2O. Samples can be assayed without any prior treatment. Some MRI contrast agents can cause transient interference in this assay.
3. Additions: A) Add 90 µL of the Chromogenic Reagent to each well containing standards, samples or controls and mix gently. B) Add 60 µL of the Calcium Assay Buffer to each well and mix gently.
4. Incubate the reaction for 5-10 minutes at room temperature. Protect from light.
5. Measure the OD at 575 nm. The chromophore is unstable and will fade slightly over time, so read the standard and samples within 30 minutes.
Calculation of Results

Calculations: Correct background by subtracting the value derived from the 0 Calcium control from all sample and standard readings (Note: The background reading may be significant and must be subtracted from sample readings). Plot standard curve µg/well vs. O.D. 575 nm readings. Then apply the sample readings to the standard curve to get Calcium sample amount in the wells (Sa). The Calcium concentrations in the test samples: C = Sa/Sv (µg/µL or mg/mL ) Where: Sa is the Calcium Sample Amount (in µg) from standard curve. Sv is the Sample Volume (µL) added into the sample well. Calcium molecular weight: 40. Calcium concentration in your sample can be expressed as mg/mL, mg/dL or mM (mM/liter). 1 mg/mL = 100 mg/dL, 1 mM = 4 mg/dL.

Restrictions For Research Use only
Storage 4 °C
Expiry Date 12 months
Product cited in: Liskova, Babchenko, Varga, Kromka, Hadraba, Svindrych, Burdikova, Bacakova: "Osteogenic cell differentiation on H-terminated and O-terminated nanocrystalline diamond films." in: International journal of nanomedicine, Vol. 10, pp. 869-84, 2015 (PubMed).

Burris, Webster, Sheriff, Faroqui, Levi, Hawse, Amlal: "Estrogen directly and specifically downregulates NaPi-IIa through the activation of both estrogen receptor isoforms (ERα and ERβ) in rat kidney proximal tubule." in: American journal of physiology. Renal physiology, Vol. 308, Issue 6, pp. F522-34, 2015 (PubMed).

Krieger, Asplin, Frick, Granja, Culbertson, Ng, Grynpas, Bushinsky: "Effect of Potassium Citrate on Calcium Phosphate Stones in a Model of Hypercalciuria." in: Journal of the American Society of Nephrology : JASN, 2015 (PubMed).

Huang, Bai, Lu, Liu, Liu, Zhu: "Osteoinductive-nanoscaled silk/HA composite scaffolds for bone tissue engineering application." in: Journal of biomedical materials research. Part B, Applied biomaterials, 2014 (PubMed).

Frick, Asplin, Culbertson, Granja, Krieger, Bushinsky: "Persistence of 1,25D-induced hypercalciuria in alendronate-treated genetic hypercalciuric stone-forming rats fed a low-calcium diet." in: American journal of physiology. Renal physiology, Vol. 306, Issue 9, pp. F1081-7, 2014 (PubMed).

Frick, Asplin, Favus, Culbertson, Krieger, Bushinsky: "Increased biological response to 1,25(OH)(2)D(3) in genetic hypercalciuric stone-forming rats." in: American journal of physiology. Renal physiology, Vol. 304, Issue 6, pp. F718-26, 2013 (PubMed).

Ming, Lv, Ma, Ge, Zhen, Song, Zhou, Ma, Xian, Chen: "The prenyl group contributes to activities of phytoestrogen 8-prenynaringenin in enhancing bone formation and inhibiting bone resorption in vitro." in: Endocrinology, Vol. 154, Issue 3, pp. 1202-14, 2013 (PubMed).

Southard, Roff, Fortugno, Richie, Kaag, Chinchilli, Virtamo, Albanes, Weinstein, Wilson: "Lead, calcium uptake, and related genetic variants in association with renal cell carcinoma risk in a cohort of male Finnish smokers." in: Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology, Vol. 21, Issue 1, pp. 191-201, 2012 (PubMed).

Lesich, Kelsch, Ponichter, Dionne, Dang, Lindemann: "The calcium response of mouse sperm flagella: role of calcium ions in the regulation of dynein activity." in: Biology of reproduction, Vol. 86, Issue 4, pp. 105, 2012 (PubMed).

Dong, Stallmann-Jorgensen, Pollock, Harris, Keeton, Huang, Li, Bassali, Guo, Thomas, Pierce, White, Holick, Zhu: "A 16-week randomized clinical trial of 2000 international units daily vitamin D3 supplementation in black youth: 25-hydroxyvitamin D, adiposity, and arterial stiffness." in: The Journal of clinical endocrinology and metabolism, Vol. 95, Issue 10, pp. 4584-91, 2010 (PubMed).

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