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Home / Assay Kits

MicroMolar Calcium / Magnesium Assay Kit

SKU : BHE16300277

This product includes 200 ul of 100 x C56 dye and 1000 ul of 10 x Reagent E and 50 ul of CaCl2. It is for measurement of 200 samples using 96-well plates.

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Mfr.No.	DMA200
Description	Calcium (Ca2+) or Magnesium (Mg2+) is an essential metal ion in biological systems. These divalent ions are also common components in biochemical buffers and pharmaceutical products. The MicoMolar Calcium / Magnesium Assay Kit is for measurement of micromolar concentrations of calcium, and magnesium (10 µM – 120 µM). The assay may also be used to detect other divalent metal ions. The assay is based on the complex formation of the divalent metal ion with the assay reagent that results in reduction of the fluorescence intensity (emission 535 nm, excitation 485 nm). Chelators such EDTA and thiol compounds bind divalent ions and should be avoided in the assay. The assay is compatible with HEPES buffer, low concentrations of non-ionic detergent (<0.01%), Tris-HCl (<10 mM), and phosphate (< 1 mM). It is not compatible with thiol compounds such as DTT, 2-mercaptoethanol or cysteine. It is interfered by divalent metal ions such as Mn2+, Fe2+, Co2+ , Ni2+ , Cu2+, Hg2+ and Zn2+. For specific assays of transitional metal ions such as Cu2+ and Zn2+ The assay kit can be used for measurements calcium or magnesium concentrations in buffer samples or pharmaceutical products. The MicroMolar Calcium / Magnesium Assay Kit (catalog number DMA200) includes 200 µl of 100 x C56 dye and 1000 µl of 10 x Reagent E and 50 µl of CaCl2. It is for measurement of 200 samples using 96-well plates. Cuvettes may also be used for measurements.
Concentration measurement	Metal ions

Product Manual	Datasheet
Protocols Description	ASSAY PROTOCOL The following assay protocol is based on using a 96-well plate for the measurement. The sample volume is 100 µl and the final assay volume is 250 µl. For assays using cuvette, the sample volume is 400 µl and the final assay volume is 1000 µl. STANDARD CURVE 1. Sample preparation: Prepare 100 µl of CaCl2 solutions in the wells of a black 96-well plate with a two-fold serial dilution from 0.2 mM to zero in water or a 10 mM HEPES, pH 7.4 buffer. For 10 samples, dilute 0.011 ml of the 100 x C56 dye 100-fold with water to make 1.1 ml of 1 x C56 dye and dilute 0.055 ml of 10 x Reagent E with water to make 0.55 ml of 1 x Reagent E. 2. Detection: Mix 50 µl of 1 x Reagent E with 100 µl of the CaCl2 solutions for 2 min. Then add 100 µl of 1 x C56 dye and incubate the solution in the dark for 15 min. Finally read the fluorescence at 535 nm (excitation at 485 nm). 3. Data Analysis: Plot the fluorescence intensity Fc and the calcium concentration [Ca] to generate the linear standard curve. Fc = a [Ca] + b Where the Fc values are from experimental data, the a and b values are from the linear fitting between the Fc values and the calcium concentrations. UNKNOWN SAMPLES Follow the same procedure to measure the fluorescence intensity Fc values from the unknown samples. Calculate the calcium concentrations in the unknown samples using the Fc values from the unknown samples and the a and b values from the standard curve. [Ca] = (Fc – b) / a

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SKU	Supplier No.	Size	Your price	Quantity
BHE16300277	DMA200	200 assays	$302.17		Add to Cart

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