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ELK Biotechnology
SKU:ELK2631
Human Alk-Smase(Alkaline Sphingomyelinase) ELISA Kit
Human Alk-Smase(Alkaline Sphingomyelinase) ELISA Kit
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Human Alk-Smase(Alkaline Sphingomyelinase) ELISA Kit
Overview
| Product name: | Human Alk-Smase(Alkaline Sphingomyelinase) ELISA Kit |
| Reactivity: | Human |
| Alternative Names: | ENPP7; E-NPP 7; NPP-7; Ectonucleotide Pyrophosphatase/Phosphodiesterase 7; Sphingomyelin Phosphodiesterase; Intestinal alkaline sphingomyelinase |
| Assay Type: | Sandwich |
| Sensitivity: | 0.113 ng/mL |
| Standard: | 20 ng/mL |
| Range: | 0.32-20 ng/mL |
| Sample Type: | tissue homogenates, cell lysates and other biological fluids |
| Assay Length: | 3.5h |
| Research Area: | Enzyme & Kinase;Neuro science; |
| Test principle: | The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Human Alk-Smase. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human Alk-Smase. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Human Alk-Smase, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Human Alk-Smase in the samples is then determined by comparing the OD of the samples to the standard curve. |
Standard curve
| Concentration (ng/mL) | OD | Corrected OD |
|---|---|---|
| 20.00 | 2.096 | 2.014 |
| 10.00 | 1.602 | 1.520 |
| 5.00 | 1.067 | 0.985 |
| 2.50 | 0.872 | 0.790 |
| 1.25 | 0.572 | 0.490 |
| 0.63 | 0.302 | 0.220 |
| 0.32 | 0.165 | 0.083 |
| 0.00 | 0.082 | 0.000 |

Precision
Intra-assay Precision (Precision within an assay):CV%<8%
Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-assay Precision (Precision between assays):CV%<10%
Three samples of known concentration were tested in forty separate assays to assess inter-assay precision.
Recovery
Matrices listed below were spiked with certain level of recombinant Alk-Smase and the recovery rates were calculated by comparing the measured value to the expected amount of Alk-Smase in samples.
| Matrix | Recovery range | Average |
|---|---|---|
| serum(n=5) | 84-106% | 100% |
| EDTA plasma(n=5) | 87-93% | 90% |
| Heparin plasma(n=5) | 80-90% | 85% |
Linearity
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Alk-Smase and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
| Matrix | 1:2 | 1:4 | 1:8 | 1:16 |
|---|---|---|---|---|
| serum(n=5) | 78-92% | 96-105% | 86-97% | 85-101% |
| EDTA plasma(n=5) | 85-94% | 87-96% | 92-105% | 89-102% |
| Heparin plasma(n=5) | 85-103% | 83-96% | 95-102% | 87-98% |
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