Phosphate Colorimetric Assay Kit(Malachite Green Method)

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Phosphate Colorimetric Assay Kit(Malachite Green Method)

SKU #	E-BC-K832-M
Detection Instrument	Microplate reader (630-650 nm)

Product Details

Properties

Sample type	Serum, plasma, tissue and cell
Sensitivity	0.001 mmol/L
Detection range	0.001 -0.20 mmol/L
Assay type	Quantitative
Assay time	35 min
Precision	Average inter-assay CV: 8% \| Average intra-assay CV: 4%
Storage	-20℃
Valid period	12 months

Dilution of Sample

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

Sample type	Dilution factor
10% Rat liver tissue homogenate	15-25
10% Rat heart tissue homogenate	15-25
10% Rat spleen tissue homogenate	15-25
10% Rat brain tissue homogenate	15-25
10% Rat kidney tissue homogenate	15-25
10% Mouse heart tissue homogenate	15-25
10% Mouse lung tissue homogenate	15-25
10% Mouse ovarian tissue homogenate	50-70
Rat serum	8-12
Rat plasma	8-12
Human plasma	8-12
10^6 HL-40 cell	2-8

Note: The diluent is normal saline (0.9% NaCl). For the dilution of other sample types, please do pretest to confirm the dilution factor

Detection Principle

The principle is that the compounds formed by phosphoric acid and molybdate under acidic conditions will form color substances with malachite green. The depth of color is proportional to the content of phosphate. The phosphorus content can be calculated indirectly be measuring the OD value at 660 nm.

Kit Components & Storage

Item	Component	Size 1(48 T)	Size 2(96 T)	Storage
Reagent 1	Chromogenic Agent A	6 mL × 1 vial	12 mL × 1 vial	2-8℃, 12 months, shading light
Reagent 2	Chromogenic Agent B	2 mL × 1 vial	4 mL × 1 vial	2-8℃, 12 months
Reagent 3	Protein Precipitator	30 mL × 1 vial	60 mL × 1 vial	2-8℃, 12 months
Reagent 4	10 mmol/L Standard Solution	0.5 mL × 1 vial	1 mL × 1 vial	2-8℃, 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)	0.03	0.08	0.15
%CV	4.2	4.0	3.8

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)	0.03	0.08	0.15
%CV	7.9	8.3	7.8

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 98%.

	Standard 1	Standard 2	Standard 3
Expected Conc. (mmol/L)	0.016	0.077	0.13
Observed Conc. (mmol/L)	0.0	0.1	0.1
Recovery rate(%)	96	99	99

Sensitivity

The analytical sensitivity of the assay is 0.001 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	0.01	0.02	0.05	0.10	0.12	0.15	0.20
Average OD	0.142	0.177	0.231	0.391	0.639	0.769	0.915	1.147
Absoluted OD	0.000	0.036	0.090	0.250	0.498	0.628	0.774	1.006