Calcium Colorimetric Assay Kit Kit
- Detection Method
- Biochemical Assay (BCA)
- 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
- 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).
- Calcium Colorimetric Assay Kit: Measure Calcium Level in a variety of Samples such as Serum, Urine etc. Convenient & Sensitive Colorimetric Assay.
Calcium Assay Buffer
Calcium Standard (500 mM)
- Application Notes
- The kit can detect calcium concentration 0.4-100 mg/dL (0.1-25 mM).
Further details regarding sample type: Cell and tissue lysates, culture media, urine, plasma and serum, as well as many other biological fluids
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.
- For Research Use only
- 4 °C
- Expiry Date
- 12 months
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- Target Type
- 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.