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Succinate Dehydrogenase (SDH) Activity Assay Kit
SKU: E-BC-K649-M-96
Succinate Dehydrogenase (SDH) Activity Assay Kit
| SKU # | E-BC-K649-M |
| Detection Instrument | Microplate reader (590-610 nm, optimum wavelength: 600 nm) |
| Detection method | Colorimetric method |
Product Details
Properties
| Synonyms | SDH |
| Sample type | Animal tissue, cell |
| Sensitivity | 0.83 U/L |
| Detection range | 0.83-65.42 U/L |
| Detection Method | Colorimetric method |
| Assay type | Enzyme Activity |
| Assay time |
10 min |
| Precision | Average inter-assay CV: 4% | Average intra-assay CV: 2% |
| Other instruments required | Centrifuge |
| Other reagents required | Normal saline (0.9% NaCl), PBS(0.01 M，pH 7.4). |
| Storage | -20℃ |
| 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 | 1 |
| 10% Rat heart tissue homogenate | 1 |
| 10% Rat kidney tissue homogenate | 1 |
| 10% Mouse liver tissue homogenate | 1 |
| 10% Mouse heart tissue homogenate | 1 |
| 10% Mouse kidney tissue homogenate | 1 |
Note: The diluent is buffer solution B. For the dilution of other sample types, please do pretest to confirm the dilution factor
Detection Principle
SDH catalyzes the dehydrogenation of succinate to fumarate, with electron transport materials transferring electrons to 2, 6-dichlorophenol indiophenol (DCPIP). Then, the reduced DCPIP is reduced to the oxidized DCPIP, which has a characteristic absorption peak at 600 nm. Therefore, the activity of SDH can be quantified by measure the change OD value at 600 nm.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Buffer Solution A | 50 mL × 1 vial | 50 mL × 2 vials | -20℃, 12 months |
| Reagent 2 | Buffer Solution B | 15 mL × 1 vial | 30 mL × 1 vial | -20℃, 12 months |
| Reagent 3 | Inhibitor | 0.8 mL × 1 vial | 0.8 mL × 2 vials | -20℃, 12 months, shading light |
| Reagent 4 | Substrate A | 1.2 mL × 1 vial | 1.2 mL × 2 vials | -20℃, 12 months, shading light |
| Reagent 5 | Substrate B | 1.2 mL × 1 vial | 1.2 mL × 2 vials | -20℃, 12 months, shading light |
| Reagent 6 | Substrate C | 0.6 mL × 1 vial | 1.2 mL × 1 vial | -20℃, 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 rat heart 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 (U/L) | 2.60 | 23.70 | 52.00 |
| %CV | 2.5 | 1.8 | 1.7 |
Inter-assay Precision
Three rat heart tissue 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) | 2.60 | 23.70 | 52.00 |
| %CV | 3.5 | 4.2 | 4.3 |
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 105%.
| Sample 1 | Sample 2 | Sample 3 | |
| Expected Conc. (U/L) | 15 | 32.5 | 58.5 |
| Observed Conc. (U/L) | 15.6 | 34.1 | 62.0 |
| Recovery rate (%) | 104 | 105 | 106 |
Sensitivity
The analytical sensitivity of the assay is 0.83 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.