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Oxaloacetate Colorimetric Assay Kit
SKU: E-BC-K901-M-96
Oxaloacetate Colorimetric Assay Kit
| SKU # | E-BC-K901-M |
| Detection Instrument | Microplate reader (402-422 nm, optimum wavelength: 412 nm) |
|
Detection method |
Colorimetric method |
Product Details
Properties
| Sample type | Animal tissue |
| Sensitivity | 0.017 mmol/L |
| Detection range | 0.017-1 mmol/L |
| Detection method | Colorimetric method |
| Assay type | Quantitative |
| Assay time | 15 min |
| Precision | Average inter-assay CV: 2.300% | Average intra-assay CV: 2.100% |
| Other instruments required | Normal saline (0.9% NaCl) |
| Other reagents required | Incubator, Ultrafiltration tube (50kD) |
| 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 | 1 |
| 10% Rat brain tissue homogenate | 1 |
| 10% Rat lung tissue homogenate | 1 |
| 10% Mouse liver tissue homogenate | 1 |
| 10% Mouse kidney tissue homogenate | 1 |
| 10% Mouse leg muscle tissue homogenate | 1 |
| 10% Pig heart tissue homogenate | 1 |
| 10% Human urine tissue homogenate | 1 |
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
Oxaloacetic acid (OAA), one of the intermediates in the tricarboxylic acid cycle is an important substance in carbon and nitrogen metabolism. Oxaloacetate reacts with substrate acetyl-coA under the action of enzyme. The substance produced can react with DTNB, which has the maximum absorption peak at 412 nm. The content of oxaloacetate can be determined by measuring the absorbance value.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Buffer Solution | 10 mL × 1 vial | 20 mL × 1 vial | -20℃, 12 months |
| Reagent 2 | Substrate | 1 mL × 1 vial | 2 mL × 1 vial | -20℃, 12 months, shading light |
| Reagent 3 | Enzyme Reagent | Liquid × 1 vial | Liquid × 2 vials | -20℃, 12 months, shading light |
| Reagent 4 | Chromogenic Agent | 1.2 mL × 1 vial | 2.4 mL × 1 vial | -20℃, 12 months, shading light |
| Reagent 5 | Standard | Power × 1 vial | Power × 2 vials | -20℃, 12 months, shading light |
| 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 rat liver 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) | 0.06 | 0.24 | 0.75 |
| %CV | 2.5 | 2.2 | 1.6 |
Inter-assay Precision
Three rat liver 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) | 0.06 | 0.24 | 0.75 |
| %CV | 2.4 | 2.2 | 2.3 |
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 101%.
| Standard 1 | Standard 2 | Standard 3 | |
| Expected Conc. (mmol/L) | 0.15 | 0.45 | 0.7 |
| Observed Conc. (mmol/L) | 0.2 | 0.4 | 0.7 |
| Recovery rate(%) | 101 | 99 | 103 |
Sensitivity
The analytical sensitivity of the assay is 0.017 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 | 0.1 | 0.2 | 0.3 | 0.5 | 0.6 | 0.8 | 1.0 |
| Average OD | 0.278 | 0.333 | 0.405 | 0.470 | 0.610 | 0.680 | 0.813 | 0.945 |
| Absoluted OD | 0 | 0.055 | 0.127 | 0.192 | 0.332 | 0.402 | 0.535 | 0.667 |