Calcium (Ca) Colorimetric Assay Kit

Calcium (Ca) Colorimetric Assay Kit

SKU #	E-BC-K103-M
Detection Instrument	Microplate reader (600-620 nm, optimum wavelength: 610 nm)
Detection Method	Colorimetric method

Product Details

Properties

Synonyms	Ca
Sample Type	Serum, plasma, urine, tissue
Sensitivity	0.07 mmol/L
Detection Range	1.5-150 mmol/L
Detection Method	Colorimetric method
Assay type	Quantitative
Assay time	20 min
Precision	Average inter-assay CV: 8.500% | Average intra-assay CV: 4.700%
Other instruments required	Micropipettor, Centrifuge, Incubator, Vortex mixer
Storage	2-8℃
Valid period	12 months

Images

TH Huang et al developed a surface decorated peptide nanoparticle that can be used for osteoporosis and diabetes therapy. Calcium (Ca) level in rat plasma was determined using Ca colorimetric assay kit (E-BC-K103-M).

 

It is recommended to take 2~3 samples with expected large difference to do pre-experiment before formal experiment and dilute the sample according to the result of the pre-experiment and the detection range (0.07-1.2 mmol/L).

The recommended dilution factor for different samples is as follows (for reference only):

Sample type	Dilution factor
Dog serum	2-3
Human serum	3-6
Mouse serum	3-6
Human urine	4-8
20% Animal tissue homogenate	1

 

Note: The diluent is deionized water.

 

Detection Principle

Calcium ion in the sample bind to Methyl Thymol Blue (MTB) in alkaline solution and form blue complex. The blue complex has a specific absorption peak at 715nm and calcium content can be calculated by measuring the OD value at 610 nm.

 

Kit Components & Storage

Item	Component	Size 1(48 T)	Size 2(96 T)	Storage
Reagent 1	MTB Reagent	5 mL ×1 vial	10 mL ×1 vial	2-8°C, 12 months, shading light
Reagent 2	Alkali Reagent	10 mL ×1 vial	20 mL × 1 vial	2-8°C, 12 months
Reagent 3	Clarificant	1 mL ×1 vial	1 mL ×1 vial	2-8°C, 12 months
Reagent 4	2.5 mmol/L Calcium Standard	5 mL ×1 vial	10 mL ×1 vial	2-8°C, 12 months
	Microplate	96 wells	No requirement
	Plate Sealer	2 pieces

 

Note: The reagents must be stored strictly according to the preservation conditions in the above table. The reagents in different kits cannot be mixed with each other. For a small volume of reagents, please centrifuge before use, so as not to obtain sufficient amount of reagents.

 

Technical Data:

Parameter:

Intra-assay Precision

Three human serum samples were assayed in replicates of 20 to determine precision within an assay. (CV = Coefficient of Variation)

Parameters	Sample 1	Sample 2	Sample 3
Mean (mmol/L)	0.35	0.86	1.05
%CV	4.8	4.7	4.6

Inter-assay Precision

Three human serum samples were assayed 20 times in duplicate by three operators to determine precision between assays.

Parameters	Sample 1	Sample 2	Sample 3
Mean (mmol/L)	0.35	0.86	1.05
%CV	8.4	8.6	8.5

 

Recovery

Take three samples of high concentration, middle concentration and low concentration to test the samples of each concentration for 6 times parallelly to get the average recovery rate of 99%

	Standard 1	Standard 2	Standard 3
Expected Conc. (mmol/L)	0.25	0.5	0.95
Observed Conc. (mmol/L)	0.3	0.5	0.9
Recovery rate (%)	101	99	97

Sensitivity

The analytical sensitivity of the assay is 0.07 mmol/L. This was determined by adding two standard deviations to the mean O.D. obtained when the zero standard was assayed 20 times, and calculating the corresponding concentration.

 

Standard Curve

As the OD value of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique or temperature effects), so the standard curve and data are provided as below for reference only:

Concentration (mmol/L)	0	0.2	0.3	0.4	0.6	0.8	1.0	1.2
OD value	0.225	0.246	0.265	0.273	0.297	0.321	0.348	0.365
0.227	0.242	0.255	0.275	0.297	0.323	0.344	0.369
Average OD	0.226	0.244	0.260	0.274	0.297	0.322	0.346	0.367
Absoluted OD	0	0.018	0.034	0.047	0.071	0.096	0.120	0.141