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Alkaline Phosphatase (ALP) Activity Assay Kit (PNPP Method)
SKU: E-BC-K009-M-500
Alkaline Phosphatase (ALP) Activity Assay Kit (PNPP Method)
| SKU # | E-BC-K009-M |
| Detection Instrument | Microplate reader(405 nm-415 nm, optimum wavelength: 405 nm) |
| Detection Method | Colorimetric method |
Product Details
Properties
| Synonyms | ALP |
| Sample Type | Serum, plasma, animal tissue, cells |
| Sensitivity | 0.27 U/L |
| Detection Range | 0.27-50.8 U/L |
| Detection Method | Colorimetric method |
| Assay Time | 30 min |
| Precision | Inter-assay CV: 4.6% | intra-assay CV: 1.2% |
| Other Reagents Required | Normal saline (0.9% NaCl) |
| 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 |
|---|---|
| Human serum | 10-20 |
| Mouse serum | 20-30 |
| Rat serum | 10-20 |
| Mouse plasma | 10-20 |
| HepG2 supernatant | 8-12 |
| 10% Rat kidney tissue homogenate | 400-1200 |
| 10% Mouse liver tissue homogenate | 10-15 |
| 10% Rat lung tissue homogenate | 50-100 |
| 10% Mouse brain tissue homogenate | 30-50 |
| Human urine | 1 |
The diluent is buffer solution. For the dilution of other sample types, please do pretest to confirm the dilution factor.
Detection Principle
Alkaline phosphatase (ALP) is a group of cytomembrane-related enzymes with hydrolysis and transfer activity, acting on a variety of phosphate substrates. ALP is a homologous dimerase and each catalytic site contains three metal ions. There are four isozymes in humans: tissue nonspecific ALP, intestinal ALP, placental ALP and genital cell ALP.
Kit Components & Storage
| Item | Component | Size 1(96 T) | Size 2 (500 Assays) | Storage |
| Reagent 1 | Buffer Solution | 60 mL×1 vial | 60 mL×5 vials | -20℃, 12 months |
| Reagent 2 | Substrate | Powder×2 vials | Powder×10 vials | -20℃, 12 months, shading light |
| Reagent 3 | 10 mmol/L p-Nitrophenol Standard Solution | 0.4 mL×1 vial | 2 mL×1 vial | 20℃, 12 months shading light |
| Reagent 4 | Stop Solution | 12 mL×1 vial | 60 mL×1 vial | -20℃, 12 month |
| UV 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) |
0.80 | 12.00 | 35.00 |
| %CV | 1.5 | 1.0 | 1.1 |
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) |
0.80 | 12.00 | 35.00 |
| %CV | 4.3 | 4.8 | 4.7 |
Sensitivity
The analytical sensitivity of the assay is 0.27 U/L ALP. 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 (μmol/L) | 0 | 40 | 80 | 160 | 240 | 320 | 400 | 500 |
| Average OD | 0.050 | 0.159 | 0.267 | 0.474 | 0.684 | 0.902 | 1.095 | 1.374 |
| Absoluted OD | 0.000 | 0.110 | 0.217 | 0.424 | 0.634 | 0.852 | 1.045 | 1.324 |