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Rat Alk-Smase(Alkaline Sphingomyelinase) ELISA Kit
ELK2629
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48 T $320.00
96 T $458.00
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Overview

Product name: Rat Alk-Smase(Alkaline Sphingomyelinase) ELISA Kit
Reactivity: Rat
Alternative Names: ENPP7; E-NPP 7; NPP-7; Ectonucleotide Pyrophosphatase/Phosphodiesterase 7; Sphingomyelin Phosphodiesterase; Intestinal alkaline sphingomyelinase
Assay Type: Sandwich
Sensitivity: 0.047 ng/mL
Standard: 10 ng/mL
Detection Range: 0.16-10 ng/mL
Sample Type: Tissue homogenates and other biological fluids.
Assay Length: 3.5h
Research Area: Enzyme & Kinase;Neuro science;
Uniprot ID: Q5EZ72
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 Rat Alk-Smase. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Rat 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 Rat 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 Rat 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
10.00 2.035 1.946
5.00 1.533 1.444
2.50 1.096 1.007
1.25 0.807 0.718
0.63 0.538 0.449
0.32 0.329 0.240
0.16 0.206 0.117
0.00 0.089 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) 78-95% 86%
EDTA plasma(n=5) 85-97% 91%
Heparin plasma(n=5) 78-93% 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) 78-93% 96-102% 95-103% 89-97%
EDTA plasma(n=5) 85-94% 87-96% 82-95% 93-102%
Heparin plasma(n=5) 96-103% 83-96% 87-98% 85-97%