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Pyruvic Acid Colorimetric Assay Kit
SKU: E-BC-K130-M-500
Pyruvic Acid Colorimetric Assay Kit
| SKU # | E-BC-K130-M |
| Detection Instrument | Microplate reader (480-520 nm, optimum wavelength: 505 nm) |
| Detection Method | Colorimetric method |
Product Details
Properties
| Sample Type | Serum, plasma, tissue |
| Sensitivity | 0.003 μmol/mL |
| Detection Range | 0.003-2.0 μmol/mL |
| Detection Method | Colorimetric method |
| Assay type | Quantitative |
| Assay time | 30 min |
| Precision | Average inter-assay CV: 3.600% | Average intra-assay CV: 2.300% |
| Other instruments required | Micropipettor, Centrifuge, Incubator, Vortex mixer |
| Other reagents required | Normal saline (0.9% NaCl), PBS (0.01 M, pH 7.4) |
| Storage | 2-8℃ |
| Valid period | 12 months |
Images
S J Tseng et al developed a nanoparticle for non-small cell lung cancer virotherapy. Pyruvic acid levels in mouse tumor cell culture medium was determined using pyruvic acid colorimetric assay kit (E-BC-K130-M).
The concentration of pyruvic acid was increased significantly after treated with nanoparticles.
Dilution of Sample
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 (0.003-2.0 μmol/mL).
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
| Human serum | 1 |
| Mouse serum | 1 |
| Mouse plasma | 1 |
| 10% Mouse liver tissue homogenate | 1 |
| 10% Rat kidney tissue homogenate | 1 |
| 10% Rat heart tissue homogenate | 1 |
Note: The diluent is normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4).
Detection Principle
Pyruvic acid can react with chromogenic agent and the reaction product is reddish brown in alkaline solution. The depth of color is directly proportional to the pyruvate content. The pyruvate content can be calculated by measuring the OD value at 505 nm.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Clarificant | 0.6 mL ×1 vial | 1.2 mL ×1 vial | 2-8°C, 12 months |
| Reagent 2 | Chromogenic Agent | 3 mL ×1 vial | 6 mL ×1 vial | 2-8°C, 12 months, shading light |
| Reagent 3 | Alkali Reagent | 10 mL ×1 vial | 20 mL ×1 vial | 2-8°C, 12 months |
| Reagent 4 | 2 μmol/mL Sodium Pyruvate Standard | 1.6 mL ×1 vial | 1.6 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 (μmol/mL) | 0.55 | 1.20 | 1.60 |
| %CV | 2.6 | 2.1 | 2.2 |
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 (μmol/mL) | 0.55 | 1.20 | 1.60 |
| %CV | 3.6 | 3.5 | 3.7 |
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 95%.
| Standard 1 | Standard 2 | Standard 3 | |
| Expected Conc. (μmol/mL) | 0.15 | 0.7 | 1.4 |
| Observed Conc. (μmol/mL) | 0.1 | 0.7 | 1.3 |
| Recovery rate (%) | 97 | 96 | 92 |
Sensitivity
The analytical sensitivity of the assay is 0.003 μmol/mL. 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 (μmol/L) | 0.0 | 0.1 | 0.2 | 0.6 | 0.8 | 1.2 | 1.6 | 2.0 |
| OD value | 0.047 | 0.105 | 0.145 | 0.326 | 0.419 | 0.601 | 0.768 | 0.896 |
| 0.049 | 0.097 | 0.152 | 0.317 | 0.415 | 0.600 | 0.769 | 0.949 | |
| Average OD | 0.048 | 0.101 | 0.148 | 0.321 | 0.417 | 0.601 | 0.768 | 0.922 |
| Absoluted OD | 0.000 | 0.053 | 0.100 | 0.273 | 0.369 | 0.553 | 0.720 | 0.874 |