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PIP3(Phosphatidylinositol Trisphosphate) ELISA Kit
ELK8250
Size： Price：
48 T $265.00
96 T $376.00
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Overview

Product name: PIP3(Phosphatidylinositol Trisphosphate) ELISA Kit
Reactivity: General
Alternative Names: PtdIns(3,4,5)P3
Assay Type: Competitive Inhibition
Sensitivity: 2.83 ng/mL
Standard: 500 ng/mL
Detection Range: 7.82-500 ng/mL
Sample Type: serum, plasma and other biological fluids
Assay Length: 2h
Research Area: Signal transduction;
Test principle: This assay employs the competitive inhibition enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with PIP3 protein. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to PIP3. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added. 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 PIP3 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
500.00 0.179
250.00 0.396
125.00 0.551
62.50 0.857
31.25 1.112
15.63 1.457
7.82 1.793
0.00 2.226

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 PIP3 and the recovery rates were calculated by comparing the measured value to the expected amount of PIP3 in samples.
Matrix Recovery range Average
serum(n=5) 82-94% 88%
EDTA plasma(n=5) 82-94% 88%
Heparin plasma(n=5) 85-97% 85%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of PIP3 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) 88-97% 91-103% 88-96% 89-97%
EDTA plasma(n=5) 85-94% 87-103% 82-94% 78-90%
Heparin plasma(n=5) 85-94% 97-105% 82-95% 89-97%