Total Price: $0.00
Contact Information
α-Ketoglutarate (α-KG) Fluorometric Assay Kit
SKU: E-BC-F047-96
α-Ketoglutarate (α-KG) Fluorometric Assay Kit
| SKU # | E-BC-F047 |
| Detection Instrument | Fluorescence microplate reader (Ex/Em=535 nm/587 nm) |
| Detection Method | Fluorescence method |
Product Details
Properties
| Synonyms | α-KG |
| Sample Type | Serum, plasma, urine, animal tissue, cell sample |
| Sensitivity | 0.60 μmol/L |
| Detection Range | 0.60-100 μmol/L |
| Detection Method | Fluorescence method |
| Assay Time | 35 min |
| Precision | Inter-assay CV: 4% | intra-assay CV: 2% |
| Other Reagents Required | Normal saline (0.9% NaCl) |
| Storage | This product can be stored at -20°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 |
|---|---|
| 10% Rat kidney tissue homogenate | 1 |
| 10% Rat lung tissue homogenate | 1 |
| 10% Porcine heart tissue homogenate | 1 |
| 10% Rat brain tissue homogenate | 1 |
| 10% Mouse liver tissue homogenate | 1 |
| 10^6 Jurkat cell | 1 |
| Human serum | 1 |
| Rat serum | 2-3 |
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
α-Ketoglutaric acid and alanine can be combined with fluorescent probe products under the action of a series of enzymes. The content of α-ketoglutaric acid in samples can be determined by measuring the fluorescence value.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size (96 T) | Storage |
| Reagent 1 | Buffer Solution | 13 mL ×1 vial | 26 mL ×1 vial | -20℃, 12 months |
| Reagent 2 | Substrate | Powder × 1 vial | Powder × 2 vials | -20℃, 12 months |
| Reagent 3 | Enzyme Reagent | Powder × 1 vial | Powder × 2 vials | -20℃, 12 months shading light |
| Reagent 4 | Probe | 1.2 mL × 1 vial | 2.4 mL × 1 vial | -20℃, 12 months shading light |
| Reagent 5 | Standard | Powder × 2 vials | Powder × 2 vials | -20℃, 12 months shading light |
| Black 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 (μmol/L) | 1.00 | 25.00 | 50.00 |
| %CV | 2.3 | 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 (μmol/L) | 1.00 | 25.00 | 50.00 |
| %CV | 4.5 | 4.1 | 3.4 |
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. (μmol/L) | 15 | 45 | 70 |
| Observed Conc. (μmol/L) | 15.2 | 44.6 | 67.9 |
| Recovery rate (%) | 101 | 99 | 97 |
Sensitivity
The analytical sensitivity of the assay is 0.60 μmol/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 fluorescence 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 | 30 | 50 | 60 | 80 | 100 |
| Fluorescence value | 547 | 1406 | 2078 | 3267 | 5050 | 6140 | 7629 | 9415 |
| 568 | 1369 | 2406 | 3370 | 5271 | 6145 | 8164 | 9746 | |
| Average fluorescence value | 557 | 1388 | 2242 | 3318 | 5161 | 6143 | 7897 | 9581 |
| Absoluted fluorescence value | 0 | 830 | 1685 | 2761 | 4603 | 5585 | 7339 | 9023 |