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Phosphate Colorimetric Assay Kit(Malachite Green Method)
SKU: E-BC-K832-M-96
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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 |