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α-L-Fucosidase (AFU) Acitivity Assay Kit
SKU: E-BC-K762-M-96
α-L-Fucosidase (AFU) Acitivity Assay Kit
| SKU # | E-BC-K762-M |
| Detection Instrument | Microplate reader (400-420 nm, optimum wavelength: 405 nm) |
Product Details
Properties
| Synonyms | AFU |
| Sample type | Serum (plasma), animal tissue |
| Sensitivity | 0.5 U/L |
| Detection range | 0.5 - 80 U/L |
| Assay type | Enzyme Activity |
| Assay time | 70 min |
| Precision | Average inter-assay CV: 8.400% | Average intra-assay CV: 3.700% |
| Other instruments required | Centrifuge, 37℃ incubator |
| Storage | 2-8℃ |
| 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 serum | 1 |
| Rat plasma | 1 |
| 10% Rat liver tissue homogenate | 1-3 |
| 10% Mouse liver tissue homogenate | 1-3 |
| 10% Rat spleen tissue homogenate | 1 |
| 10% Mouse kidney tissue homogenate | 1 |
Note: The diluent is extraction solution. For the dilution of other sample types, please do pretest to confirm the dilution factor.
Detection Principle
α-L-fucosidase (AFU) is an acid hydrolase that exists in organisms and widely exists in tissues, cells, blood and body fluids. AFU is involved in the metabolism of glycoproteins, glycolipids and oligosaccharides, and plays a key role in cell differentiation, apoptosis, inflammation and host-pathogen interaction. AFU catalyzes the hydrolysis of the colorless substrate nitrophenyl-α-L-fucoside to produce yellow nitrophenol with a significant absorption peak at 405 nm, and its absorbance is positively correlated with the content of nitrophenol. The activity of AFU can be calculated by measuring the change of absorbance value at 405 nm.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Extraction Solution | 50 mL × 1 vial | 50 mL × 2 vials | 2-8℃, 12 months |
| Reagent 2 | Working Solution | 12 mL × 1 vial | 25 mL × 1 vial | 2-8 ℃, 12 months, shading light |
| Reagent 3 | 5 mmol/L Standard Solution | 1 mL × 1 vial | 1 mL × 2 vials | 2-8 ℃, 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 (U/L) | 3.50 | 34.60 | 72.00 |
| %CV | 4.0 | 3.6 | 3.5 |
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 (U/L) | 3.50 | 34.60 | 72.00 |
| %CV | 8.2 | 8.5 | 8.5 |
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 97%.
| Standard 1 | Standard 2 | Standard 3 | |
| Expected Conc. (mmol/L) | 0.7 | 1.8 | 3.3 |
| Observed Conc. (mmol/L) | 0.7 | 1.7 | 3.2 |
| Recovery rate (%) | 97 | 96 | 98 |
Sensitivity
The analytical sensitivity of the assay is 0.5 U/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 | 0.5 | 1 | 1.5 | 2 | 3 | 4 | 5 |
| Average OD | 0.093 | 0.269 | 0.410 | 0.590 | 0.739 | 1.073 | 1.407 | 1.731 |
| Absoluted OD | 0.000 | 0.176 | 0.317 | 0.497 | 0.646 | 0.980 | 1.314 | 1.638 |