Calcium Colorimetric Assay Kit

K380 is available from Abcam as ab102505.
Catalog #: K380 | abID: ab102505

Product Details

abID ab102505
Cat # +Size K380-250
Size 250 assays
Detection Method Absorbance (575 nm)
Species Reactivity Mammalian
Applications The kit can detect calcium concentration 0.4-100 mg/dL (0.1-25 mM).
Features & Benefits • Simple procedure; takes less than 20 minutes
• Fast and convenient
• The assay is sensitive, stable and high-throughput adaptable
Kit Components • Calcium Assay Buffer
• Chromogenic Reagent
• Calcium Standard (500 mM)
Storage Conditions 4°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.


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. BioVision’s 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).

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Does the kit measure free Ca2+ ions or total calcium (free and bound)?
The kit utilizes the chromogenic complex (λ = 575 nm) formed between calcium ions and 0-cresolphthalein. Therefore, it measures only free calcium and not the bound calcium.
Which anticoagulants DO NOT interfere with Calcium assay?
Heparin is the ONLY anticoagulant that DOES NOT interfere
What about hemoglobin/hemolysis?
Assay will work with = 750 mg/ml hemaglobin
A customer is interested in the calcium assay kit, K380-250. The protocol recommends using 2x10^6 cells in 500 uL of buffer, however she will only have 50,000-150,000 cells per sample. Each test only requires 2-50 uL of samples however. Could the sample preparation be scaled down, i.e. use 50 uL of buffer with 2x10^5 cells?
Yes, surely it can be scaled down ?. My only concern is that if she is making samples in just 50 µl, she will not have enough for the pilot expt and duplicates for the main expt. Maybe she can have more cells and volume for just one sample to do the pilot expt and determine if 50 µl homogenate is enough for duplicates in the main expt. But in a nutshell, you can scale down the volumes to suit your needs as long as you still do the final expt properly.
I want to meassure the CA+2 in neutrophils. You have a specific buffer for their preparation (lysis)?
You can use the Calcium assay buffer provided with the kit. Use 2-5 x 10^6 cells, homogenize with the buffer, centrifuge and take the sup for the assay.
The customer wants to use K380-250 to measure the calcium contained in cell culture samples and has no access to a sonicator. What would you recommend for the sample preparation?
They can use a dounce homogenizer. Alternatively freeze/thaw cycles with vortexing in the middle can also lyse the cells, although I would go with that as the last option.
Can this kit be used with samples like bacteria, plants, drosophila, yeast etc?
We have optimized the kit with mammalian samples. However, theoretically these kits should work with samples from multiple species/sources. Since the optimal conditions depend on the sample type, the protocol has to be be adapted to fit the samples for efficient results. Please refer to this kit's citations to see what kind of samples have been used with this kit other than mammalian samples.
Can we use frozen samples with this assay?
Fresh samples are always preferred over frozen samples. However, frozen samples can also be used, provided, they were frozen right after isolation, were not freeze thawed multiple time (for which we recommend aliquoting the samples before freezing) and have been frozen for relatively short periods.
Can we use a different wavelength than recommended for the final analysis?
It is always recommended to use the exact recommended wavelength for the most efficient results. However, most plate readers have flexibility in their band width of detection in increments of +/- 10 nm. Depending on this flexibility range, you can deviate from the recommended wavelengths within limits.
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. You have to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Do you have trial sizes of this kit?
Unfortunately, we do not have trial sizes of this kit available. However, if you are based in the US or Canada, we will give you a 10% off list price introductory discount on its purchase price. If you are based out of this area please contact your regional BioVision distributor.
What is the shelf life of this kit?
This kit is good for 12 months from the date of shipment in the unopened form when stored at the appropriate temperature and appropriate conditions. After opening and reconstitution, some of the components in this kit are good for 2 months at -20°C. Please refer to the datasheet for storage information and shelf life of each of the components.
Why are my standard curve values lower than those shown on the datasheet?
There are multiple factors which influence the signals like the incubation times, room temperature, handling etc. In general, to increase the value of the standards, you can increase the incubation time. As long as the standard curve is linear, it should be fine to use, since all of your samples will also be measured under the same conditions on this curve.
How do I normalize my samples against protein concentration
You can use a protein quantitation assay on the supernatants you get from cell/tissue lysates or with any other liquid sample in the assay buffer.
Can we purchase individual components of this kit?
Yes, you can purchase any of the kit's components without the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can we use an alternate buffer for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.
Should I make a standard curve for every expt I do, or is one curve/kit enough?
Yes, I would strongly recommend you to do the standards every time you do the expt. There is always a chance that something was done differently that day and we do not want any conditions to differ between standards and samples.
Jiao Jiao Zhang, et al., (2018) MicroRNA-7450 regulates non-thermal plasma-induced chicken Sertoli cell apoptosis via adenosine monophosphate-activated protein kinase activation, Scientific Reports, Jun. 2018, 29884805
Alenzi, Mohammed et al. (2017) Antiurolithic effect of olive oil in a mouse model of ethylene glycol-induced urolithiasis, Investig Clin Urol. 2017 May;58(3):210-216.
Wei Wei et al. (2017) In vitro osteogenic induction of bone marrow mesenchymal stem cells with a decellularized matrix derived from human adipose stem cells and in vivo implantation for bone regeneration, J. Mater. Chem. B, 2017,5, 2468-2482
Chang et al., Effect of ED-71, an analogue of Vitamin D3, on intestinal neoplasia in the Apc+/Min-FCCC mouse model. Cancer Res., Aug 2015; 75: 2806.
Krieger et al., Effect of Potassium Citrate on Calcium Phosphate Stones in a Model of Hypercalciuria. J. Am. Soc. Nephrol., Apr 2015; 10.1681/ASN.2014121223.
For more citations of this product click here