Total Price: $0.00
Contact Information
Total Amino Acids (T-AA) Colorimetric Assay Kit
SKU: E-BC-K055-M-96
To better serve you, we would like to discuss your specific requirement.
Please Contact Us for a quote.
Total Amino Acids (T-AA) Colorimetric Assay Kit
| SKU # | E-BC-K055-M |
| Detection Instrument | Microplate reader (640-660 nm, optimum wavelength: 650 nm) |
| Detection Method | Colorimetric method |
Product Details
Properties
| Synonyms | T-AA |
| Sample Type | Serum, plasma, urine, tissue |
| Sensitivity | 3.03 mmol/L |
| Detection Range | 3.64-100 mmol/L |
| Detection Method | Colorimetric method |
| Assay type | Quantitative |
| Assay time | 40 min |
| Precision | Average inter-assay CV: 6.500% | Average intra-assay CV: 4% |
| Other instruments required | Test tube, Micropipettor, Vortex mixer, Centrifuge |
| Other reagents required | Normal saline (0.9% NaCl), PBS (0.01 M, pH 7.4) |
| Storage | 2-8℃ |
| Valid period | 12 months |
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 (3.64-100 mmol/L).
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
| Human serum | 1 |
| Human urine | 1 |
| Rat plasma | 1 |
| Porcine serum | 1 |
| 10% Rat heart tissue homogenate | 1 |
| 10% Rat liver tissue homogenate | 1 |
| 10% Mouse liver homogenate | 1 |
| 10% Epipremnum aureum leaf tissue homogenate | 1 |
Note: The diluent is reagent 5.
Detection Principle
Copper ions can complex with various amino acids to produce blue-green complex compound, and the depth of color is proportional to the content of total amino acids at a specific wavelength. T-AA content can be calculated with the absorbance at 650 nm.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Powder A | Powder × 1 vial | Powder × 1 vial | 2-8℃, 12 months |
| Reagent 2 | Acid Reagent | 0.4 mL × 1 vial | 0.8 mL × 1 vial | 2-8℃, 12 months |
| Reagent 3 | Powder B | Powder × 1 vial | Powder × 1 vial | 2-8°C, 12 months |
| Reagent 4 | Powder C | Powder × 1 vial | Powder × 1 vial | 2-8℃, 12 months |
| Reagent 5 | Protein Precipitator | 8 mL ×1 vial | 15 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) | 10.50 | 42.60 | 88.10 |
| %CV | 4.2 | 3.8 | 4.0 |
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) | 10.50 | 42.60 | 88.10 |
| %CV | 7.0 | 6.3 | 6.2 |
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 102%.
| Standard 1 | Standard 2 | Standard 3 | |
| Expected Conc. (mmol/L) | 14.5 | 45 | 73.2 |
| Observed Conc. (mmol/L) | 14.6 | 44.6 | 77.6 |
| Recovery rate (%) | 101 | 99 | 106 |
Sensitivity
The analytical sensitivity of the assay is 3.03 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 (μmol/L) | 0 | 10 | 20 | 40 | 50 | 60 | 80 | 100 |
| Average OD | 0.070 | 0.086 | 0.098 | 0.126 | 0.141 | 0.156 | 0.186 | 0.214 |
| Absoluted OD | 0.000 | 0.016 | 0.028 | 0.056 | 0.071 | 0.086 | 0.116 | 0.144 |