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Malic Acid Colorimetric Assay Kit
SKU: E-BC-K905-M-96
Malic Acid Colorimetric Assay Kit
| SKU # | E-BC-K905-M |
| Detection Instrument | Microplate reader (445-465 nm, optimum wavelength: 450 nm) |
| Detection method: | Colorimetric method |
Product Details
Properties
| Sample type | Serum (plasma), tissue and cell |
| Sensitivity | 0.003 mmol/L |
| Detection range | 0.003-0.5 mmol/L |
| Detection method | Colorimetric method |
| Assay type | Quantitative |
| Precision | Average inter-assay CV: 4.6-5.9% | Average intra-assay CV: 2.6-3.6% |
| 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 | 3-5 |
| 10% Mouse liver tissue homogenate | 3-5 |
| 10% Lemon tissue homogenate | 8-12 |
| 10% Tangerine tissue homogenate | 5-10 |
| 10% Potato tissue homogenate | 5-12 |
| Rat serum | 1 |
| Mouse plasma | 1 |
| 3.35×106 CHO cells | 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
L-malic acid, also known as 2-hydroxysuccinic acid, is widely present in living organisms. It has two stereoisomers and is one of the members of the tricarboxylic acid cycle, an important metabolic cycle in living organisms. The detection principle of this kit is that the enzyme catalyzes malic acid to produce oxaloacetic acid. At the same time, under the action of electronic coupler, the chromogenic agent is reduced to produce orange yellow product, which has the maximum absorption peak at about 450 nm.
Kit Components & Storage
| Item | Component | Size (96 T) | Storage |
| Reagent 1 | Buffer Solution | 20 mL × 1 vial | -20°C, 12 months |
| Reagent 2 | Enzymatic Reagent | 0.3 mL × 1 vial | -20°C, 12 months, shading light |
| Reagent 3 | Chromogenic Agent | 2.4 mL × 1 vial | -20°C, 12 months, shading light |
| Reagent 4 | Substrate | Powder × 2 vials | -20°C, 12 months, shading light |
| Reagent 5 | 0.5 mmol/L Standard | 5 mL × 1 vial | -20°C, 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 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) | 0.06 | 0.10 | 0.30 |
| %CV | 3.6 | 3.4 | 2.6 |
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) | 0.06 | 0.10 | 0.30 |
| %CV | 4.6 | 5.9 | 5.1 |
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) | 0.13 | 0.27 | 0.38 |
| Observed Conc. (mmol/L) | 0.1 | 0.3 | 0.4 |
| Recovery rate(%) | 101 | 96 | 97 |
Sensitivity
The analytical sensitivity of the assay is 0.003 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 | 0.15 | 0.2 | 0.3 | 0.35 | 0.4 | 0.5 |
| Average OD | 0.064 | 0.130 | 0.157 | 0.191 | 0.257 | 0.292 | 0.320 | 0.383 |
| Absoluted OD | 0 | 0.059 | 0.093 | 0.127 | 0.193 | 0.229 | 0.257 | 0.319 |