Sucrose Colorimetric Assay Kit
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Sucrose Colorimetric Assay Kit
| SKU # | E-BC-K161-S |
| Detection Instrument | Spectrophotometer (290 nm) |
| Detection method | Colorimetric method |
Product Details
Properties
| Sample Type | Plant tissue |
| Sensitivity | 0.32 μmol/mL |
| Detection Range | 0.32-70 μmol/mL |
| Detection Method | Colorimetric method |
| Assay type | Quantitative |
| Assay time | 40 min |
| Precision | Average inter-assay CV: 8.500% | Average intra-assay CV: 3.400% |
| Other instruments required | Micropipettor, Vortex mixer, 100℃ Water bath |
| Other reagents required | 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 (0.32-70 μmol/mL).
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
| 10% Green pepper tissue homogenization | 1 |
| 10% Epipremnum aureum tissue homogenization | 1 |
| 10% Cucumber tissue homogenization | 1 |
Note: The diluent is normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4).
Detection Principle
Sucrose in plant tissue is hydrolyzed to glucose and fructose in boiling water bath under acidic conditions. 5-hydroxymethyl furfural was synthesized from fructose under acid condition and measure the ultraviolet absorption of 5-hydroxymethyl furfural. Glucose must be dissimilated into ketose structure and reduced to obtain 5-hydroxymethylfurfural, but the rate of isomerization of glucose to ketose is very slow. Therefore, the ultraviolet absorption of glucose is much smaller than fructose.
Kit Components & Storage
| Item | Component |
Size 1 (50 Assays) |
Size 2 (100 Assays) |
Storage |
| Reagent 1 | Hydrolysate Solution | 60 mL × 2 vials | 60 mL × 4 vials | 2-8°C, 12 months |
| Reagent 2 | 100 μmol/mL Sucrose Standard | 1 mL × 1 vial | 1 mL × 1 vial | 2-8°C, 12 months |
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 cucumber tissue 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/mL) | 8.50 | 34.80 | 58.00 |
| %CV | 3.7 | 3.4 | 3.1 |
Inter-assay Precision
Three rat liver tissue samples were assayed 20 times in duplicate by three operators to determine precision between assays.
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (μmol/mL) | 8.50 | 34.80 | 58.00 |
| %CV | 8.3 | 8.1 | 9.1 |
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. (μmol/mL) | 12.6 | 45.5 | 60 |
| Observed Conc. (μmol/mL) | 13.4 | 44.1 | 61.8 |
| Recovery rate (%) | 106 | 97 | 103 |
Sensitivity
The analytical sensitivity of the assay is 0.32 μmol/mL. 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 | 10 | 20 | 30 | 40 | 50 | 70 |
| Average OD | 0.007 | 0.296 | 0.600 | 0.921 | 1.220 | 1.547 | 2.071 |
| Absoluted OD | 0 | 0.289 | 0.593 | 0.914 | 1.213 | 1.540 | 2.064 |