L-Arginine (L-Arg) Colorimetric Assay Kit
L-Arginine (L-Arg) Colorimetric Assay Kit
| SKU # | E-BC-K850-M |
| Detection Instrument | Microplate reader (510-530 nm, optimum wavelength: 520 nm) |
| Detection method: | Colorimetric method |
Product Details
Properties
| Synonyms | L-Arg |
| Sample type | Serum, plasma, urine, plant and animal tissue |
| Sensitivity | 0.008 mmol/L |
| Detection range | 0.008-1 mmol/L |
| Detection method |
Colorimetric method |
| Assay type | Quantitative |
| Precision | Average inter-assay CV: 6-8% | Average intra-assay CV: 1.8-3.8% |
| Other instruments required | 60℃ and 100℃ water bath, 10 KD ultrafiltration tube |
| Other reagents required | Normal saline (0.9% NaCl) |
| 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 | 1-2 |
| 10% Rat kidney tissue homogenate | 1-2 |
| 10% Rat heart tissue homogenate | 1-2 |
| 10% Rat lung tissue homogenate | 1-2 |
| Human serum | 1-2 |
| Human plasma | 1-2 |
| Bovine serum | 1-2 |
| 10% Nartjie tissue homogenate | 1-2 |
Note: The diluent is buffer solution A. For the dilution of other sample types, please do pretest to confirm the dilution factor.
Detection Principle
Arginine (Arg) is an essential amino acid in the human body and can also be absorbed in food. Natural arginine is L-type, which is alkaline and easily reacts with acids to form salts. Arginine is an intermediate metabolite in the ornithine cycle, which can promote the conversion of ammonia into urea, thereby reducing blood ammonia content.Arginine generates urea under the action of enzymes, and urea can complex with oxime reagents under strong acidic conditions by boiling. The complex has a maximum absorption peak at 520 nm, and the arginine content can be calculated by measuring the OD value at 450 nm.
Kit Components & Storage
| Item | Component | Size (96 T) | Storage |
| Reagent 1 | Buffer Solution A | 50 mL × 1 vial | -20℃, 12 months |
| Reagent 2 | Buffer Solution B | 1.8 mL × 1 vial | -20℃, 12 months, shading light |
| Reagent 3 | Enzyme Reagent A | Powder ×2 vials | -20℃, 12 months, shading light |
| Reagent 4 | Enzyme Reagent B | Powder ×2 vials | -20℃, 12 months, shading light |
| Reagent 5 | Chromogenic Agent A | 24 mL ×1 vial | -20℃, 12 months, shading light |
| Reagent 6 | Chromogenic Agent B | 24 mL ×1 vial | -20℃, 12 months, shading light |
| Reagent 7 | 1 mmol/L Standard | 1 mL ×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 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) | 0.05 | 0.32 | 0.74 |
| %CV | 3.8 | 1.9 | 1.8 |
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) | 0.05 | 0.32 | 0.74 |
| %CV | 8.0 | 6.0 | 7.6 |
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 96%.
| Standard 1 | Standard 2 | Standard 3 | |
| Expected Conc. (mmol/L) | 0.15 | 0.42 | 0.7 |
| Observed Conc. (mmol/L) | 0.1 | 0.4 | 0.7 |
| Recovery rate(%) | 98 | 94 | 96 |
Sensitivity
The analytical sensitivity of the assay is 0.008 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 (μg/mL) | 0 | 0.1 | 0.2 | 0.3 | 0.5 | 0.6 | 0.8 | 1.0 |
| OD Value | 0.064 | 0.115 | 0.173 | 0.224 | 0.330 | 0.370 | 0.464 | 0.579 |
| 0.064 | 0.115 | 0.173 | 0.224 | 0.328 | 0.370 | 0.471 | 0.584 | |
| Average OD | 0.064 | 0.115 | 0.173 | 0.224 | 0.329 | 0.370 | 0.468 | 0.582 |
| Absoluted OD | 0 | 0.051 | 0.109 | 0.160 | 0.265 | 0.306 | 0.404 | 0.518 |