The calcium assay is used to measure the activity of the ionized calcium in samples of whole blood. This test requires that the sample be treated within half an hour. This is because the whole blood sample is sensitive to ionized calcium and must be treated immediately. The serum obtained from the blood samples can be stored for up to six weeks in the refrigerator. The tubes used to collect the serum must be tightly sealed to ensure its quality and reliability. Several methods are available to measure ionized calcium in a laboratory setting, including a high-throughput method using a single-tube instrument.
The first steps of the calcium assay are very simple. The first step is to mix the samples in a microplate. Then, the cells are incubated for 9 days. The second step is decalcification of the cells by adding 0.6 N HCl to the cell culture medium. The supernatants are then analyzed with the QuantiChrom Calcium Assay Kit (BioAssay Systems). The difference between the two means is then determined using a statistical test. For the second step, the P value must be less than 0.5.
Steps Taken Using The Assay
The third step is to prepare the sample for the calcium assay. The control wells have already been diluted with the sample mixture and a dose of ionized calcium. During the first addition of the control, the ionomycin saturates the calcium indicators. Then, the second step is to mix the samples. The last step is to store the sample in the tube tightly. Then, the samples should be weighed and placed into the chamber.
For the calcium assay, a multimode plate reader is the most ideal choice. It has ultra-fast data sampling and the most sophisticated bottom reading optical system on the market. In addition, the reader is equipped with several measurement modes, including temperature, humidity, and pH. It also has an Atmospheric Control Unit and multiple shaking options. There are reagent injectors available for the QC-96-well format.
The calcium assay kit contains an anion-impermeable fluorescence quencher. The cho-M1 cell line expresses P2Y receptors. The CHO-M1 cells were seeded at a density of 40,000 cells per 100 ul in a 96-well microplate. After incubation, the CHO cells were incubated with a solution containing one hundred ul of the ionized calcium assay. A few minutes later, the color of the cell membrane was detected, and the resulting signal was a blue-ish signal.
Unlike the calcium assay, the Fluo-4 Direct Calcium Assay kit can be used on Fluorescence microplate readers with bottom-read mode. Its two-dimensional fluorescent dye, called Calbryte(TM), is loaded into the cell layer of interest. The result is a uniform, standardized signal. This assay kit can be used for a variety of applications, and the results can be easily interpreted in a clinical research setting.