Glycogen Fluorometric Assay Kit
Glycogen Fluorometric Assay Kit
| SKU # | E-BC-F040 |
| Detection Instrument | Fluorescence microplate reader (Ex/Em=535 nm/587 nm) |
| Detection Method | Fluorescence method |
Product Details
Properties
| Sample Type | Animal liver, muscle tissue |
| Sensitivity | 0.06 μg/mL |
| Detection Range | 0.06-4.0 μg/mL |
| Detection Method | Fluorescence method |
| Assay Time | 50 min |
| Precision | Inter-assay CV: 6.6% | intra-assay CV: 3.4% |
| 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 liver tissue homogenate | 3000-5000 |
| 10% Mouse muscle tissue homogenate | 10-20 |
The diluent is buffer solution A. For the dilution of other sample types, please do pretest to confirm the dilution factor.
Detection Principle
Glycogen produces glucose under the action of starch glycosidase, and glucose is catalyzed by glucose oxidase to produce hydrogen peroxide. In the presence of the peroxidase, hydrogen peroxide be oxidized to produce the fluorescence substrate. The fluorescence intensity at the excitation wavelength of 535 nm and emission wavelength of 587 nm is proportional to the glycogen content.
Kit Components & Storage
| Item | Component | Size (96 T) | Storage |
| Reagent 1 | Buffer Solution A | 30 mL ×1 vial | -20℃, 12 months |
| Reagent 2 | Buffer Solution B | 8 mL × 1 vial | -20℃, 12 months |
| Reagent 3 | Probe | 0.24 mL × 1 vial | -20℃, 12 months shading light |
| Reagent 4 | Enzyme Reagent A | Powder × 1 vial | -20℃, 12 months |
| Reagent 5 | Enzyme Reagent B | Powder × 1 vial | -20℃, 12 months |
| Reagent 6 |
0.1 mg/mL Glucogen Standard Solution |
0.5 mL × 1 vial | -20℃, 12 months |
| 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 rat liver 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 (μg/mL) | 0.15 | 1.00 | 2.50 |
| %CV | 4.0 | 3.2 | 3.0 |
Inter-assay Precision
Three rat liver samples were assayed 20 times in duplicate by three operators to determine precision between assays.
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (μg/mL) | 0.15 | 1.00 | 2.50 |
| %CV | 7.3 | 6.2 | 6.3 |
Sensitivity
The analytical sensitivity of the assay is 0.06 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 | 0.5 | 1.0 | 1.5 | 2.0 | 2.5 | 3.0 | 4.0 |
| Fluorescence value | 1269 | 2695 | 3650 | 5304 | 6826 | 8119 | 9197 | 11360 |
| 1257 | 2599 | 3519 | 5209 | 6885 | 7861 | 9271 | 11325 | |
| Average fluorescence value | 1263 | 2647 | 3585 | 5257 | 6856 | 7990 | 9234 | 11343 |
| Absoluted fluorescence value | 0 | 1384 | 2322 | 3994 | 5593 | 6727 | 7971 | 10080 |