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ELK Biotechnology

ELK9484

General SIP(Sphingosine 1 Phosphate) ELISA Kit

General SIP(Sphingosine 1 Phosphate) ELISA Kit

Sensitivity

General SIP(Sphingosine 1 Phosphate) ELISA Kit

Overview

Product name: SIP(Sphingosine 1 Phosphate) ELISA Kit
Reactivity: General
Alternative Names: EDG3; LPB3; S1P3; S1PR3; endothelial differentiation, sphingolipid G-protein-coupled receptor, 3; Sphingosine 1-phosphate receptor Edg-3
Assay Type: Sandwich
Sensitivity: 0.91 ng/mL
Standard: 200 ng/mL
Detection Range: 3.13-200 ng/mL
Sample Type: Tissue homogenates, cell lysates and other biological fluids
Assay Length: 3.5h
Research Area: Signal transduction;
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 SIP. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to SIP. 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 SIP, 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 SIP 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
200.00 2.135 2.052
100.00 1.638 1.555
50.00 1.173 1.090
25.00 0.871 0.788
12.50 0.525 0.442
6.25 0.357 0.274
3.13 0.252 0.169
0.00 0.083 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 SIP and the recovery rates were calculated by comparing the measured value to the expected amount of SIP in samples.
Matrix Recovery range Average
serum(n=5) 88-103% 95%
EDTA plasma(n=5) 88-102% 95%
Heparin plasma(n=5) 80-95% 87%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of SIP 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) 93-104% 84-97% 88-97% 95-106%
EDTA plasma(n=5) 83-98% 81-96% 87-105% 83-92%
Heparin plasma(n=5) 84-99% 88-103% 83-94% 92-105%
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