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D-Lactic Acid/Lactate(LA)Colorimetric Assay Kit
SKU: E-BC-K002-M-500
D-Lactic Acid/Lactate(LA)Colorimetric Assay Kit
| SKU # | E-BC-K002-M |
| Detection Instrument | Microplate reader(520 nm-550 nm, optimum wavelength: 530 nm) |
| Detection Method | Colorimetric method |
Product Details
Properties
| Synonyms |
D-LA |
| Sample Type | Serum, plasma, animal tissue, saliva sample |
| Sensitivity | 0.06 mmol/L |
| Detection Range | 0.06-8.0 mmol/L |
| Detection Method | Colorimetric method |
| Assay Time | 20 min |
| Precision | Inter-assay CV: 7.7% | intra-assay CV: 3.8% |
| Other Reagents Required | PBS (0.01 M, pH 7.4) |
| Storage | This product can be stored at 2~8°C for 12 months with shading light. |
| Valid period | 12 months |
Dilution Of Sample
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
|---|---|
| Human plasma | 1 |
| Human serum | 1 |
| Rat serum | 1 |
| Rat plasma | 1 |
| Mouse serum | 1 |
| Rabbit serum | 1 |
| 10% Rat kidney tissue homogenate | 2-3 |
| 10% Mouse brain tissue homogenate | 1 |
The diluent is normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4). For the dilution of other sample types, please do pretest to confirm the dilution factor.
Detection Principle
Using NAD+ as H+ receptor, D-lactate dehydrogenase (LDH) catalyzes the reaction of D-lactic acid and NAD+ to generate pyruvic acid and NADH respectively. NBT is reduced to a kind of purple compound during the reaction. Measure the OD value at 530 nm, and the concentration of D-lactic acid can be calculated.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Buffer Solution | 6 mL ×1 vial | 12 mL ×1 vial | 2-8℃, 12 months |
| Reagent 2 | Enzyme Stock Solution |
0.06 mL ×1 vial | 0.12 mL × 1 vial | 2-8℃, 12 months |
| Reagent 3 | Chromogenic Agent | 1.2 mL ×1 vial | 1.2 mL × 2 vials | 2-8℃, 12 months shading light |
| Reagent 4 | Stop Solution | 12 mL ×1 vial | 24 mL × 1 vial | 2-8℃, 12 months |
| Reagent 5 | 10 mmol/L Standard Solution |
1.0 mL ×1 vial | 2.0 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) |
0.50 | 2.50 | 5.00 |
| %CV | 4.2 | 3.6 | 3.6 |
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) |
0.50 | 2.50 | 5.00 |
| %CV | 8.2 | 7.5 | 7.4 |
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 99%.
| Standard 1 | Standard 2 | Standard 3 | |
| Expected Conc. (mmol/L) | 1.5 | 4.5 | 6.5 |
| Observed Conc. (mmol/L) | 1.5 | 4.4 | 6.4 |
| Recovery rate (%) | 101 | 98 | 98 |
Sensitivity
The analytical sensitivity of the assay is 0.06 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 | 1.0 | 2.0 | 4.0 | 5.0 | 6.0 | 7.0 | 8.0 |
| OD value | 0.138 | 0.284 | 0.414 | 0.648 | 0.752 | 0.856 | 0.934 | 1.083 |
| 0.136 | 0.288 | 0.405 | 0.620 | 0.765 | 0.823 | 0.961 | 1.032 | |
| Average OD | 0.137 | 0.286 | 0.410 | 0.634 | 0.759 | 0.840 | 0.948 | 1.058 |
| Absoluted OD | 0.000 | 0.149 | 0.273 | 0.497 | 0.622 | 0.703 | 0.811 | 0.921 |