Dipeptidyl Peptidase IV (DPP4) Activity Fluorometric Assay Kit
Dipeptidyl Peptidase IV (DPP4) Activity Fluorometric Assay Kit
| SKU # | E-BC-F053 |
| Detection Instrument | Fluorescence microplate reader (Ex/Em=360 nm/460 nm) |
| Detection Method | Fluorescence method |
Product Details
Properties
| Synonyms | DPP4 |
| Sample Type | Serum, plasma, tissue, cell |
| Sensitivity | 0.92 U/L |
| Detection Range | 0.92-3.77 U/L |
| Detection Method | Fluorescence method |
| Assay Time | 70 min |
| Precision | Inter-assay CV: 5.7% | intra-assay CV: 0.6% |
| 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 |
|---|---|
| 5% Mouse kidney tissue homogenate | 4-5 |
| 5% Mouse lung tissue homogenate | 4-5 |
| 5% Mouse heart tissue homogenate | 4-5 |
| 5% Rat liver tissue homogenate | 4-5 |
| 5% Rat brain tissue homogenate | 4-5 |
| Porcine serum | 4-5 |
| Horse serum | 4-5 |
| Dog serum | 4-5 |
| Female chicken serum | 4-5 |
| Macaca fascicularis serum | 4-5 |
| 5.0×10^6 HL-60 cell | 1 |
| 4.1×10^6 293T cell | 1 |
The diluent is buffer solution. For the dilution of other sample types, please do pretest to confirm the dilution factor.
Detection Principle
Dipeptidyl Peptidase IV (DPP4), also known as CD26, is a kind of serine protease, can decompose the second N-terminal proline or alanine residue of the peptide chain. In organisms, dipeptidyl peptidase can rapidly decompose incretin, which can stabilize insulin level and promote the reduction of blood glucose level in organisms. The detection principle of this kit is that DPP4 can decompose the substrate and release the fluorescent substance AMC, and the activity of this enzyme can be inhibited by adding DPP4 inhibitor. The activity of DPP4 could be calculated by the difference of fluorescence value before and after inhibition.
Kit Components & Storage
| Item | Component | Size (96 T) | Storage |
| Reagent 1 | Buffer Solution | 60 mL × 1 vial | -20℃, 12 months |
| Reagent 2 | Postive Control | Powder ×1 vial | -20℃, 12 months shading light |
| Reagent 3 | Substrate | 1.2 mL ×2 vials | -20℃, 12 months shading light |
| Reagent 4 | Inhibitors | 0.3 mL × 2 vials | -20℃, 12 months shading light |
| Reagent 5 | 1 mmol/L Standard | 1 mL × 1 vial | -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 (U/L) | 1.00 | 2.50 | 3.00 |
| %CV | 1.0 | 0.4 | 0.4 |
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) | 1.00 | 2.50 | 3.00 |
| %CV | 4.5 | 6.2 | 6.4 |
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 100%.
| Standard 1 | Standard 2 | Standard 3 | |
| Expected Conc. (μmol/L) | 25 | 55 | 75 |
| Observed Conc. (μmol/L) | 24.8 | 55.0 | 75.8 |
| Recovery rate (%) | 99 | 100 | 101 |
Sensitivity
The analytical sensitivity of the assay is 0.92 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 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 | 20 | 40 | 50 | 60 | 70 | 80 | 100 |
| Fluorescence value | 16 | 6862 | 12505 | 15781 | 18403 | 21563 | 24781 | 28544 |
| 20 | 7082 | 12876 | 16059 | 19481 | 22312 | 25503 | 29309 | |
| Average fluorescence value | 18 | 6972 | 12691 | 15920 | 18942 | 21938 | 25142 | 28927 |
| Absoluted fluorescence value | 0 | 6954 | 12673 | 15902 | 18924 | 21920 | 25124 | 28909 |