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Caffeine ELISA Kit
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ELK0683
Size： Price：
48 T $320.00
96 T $458.00
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Overview

Product name: Caffeine ELISA Kit
Reactivity: General
Alternative Names: Caffeine
Assay Type: Competitive Inhibition
Sensitivity: 0.25 ng/mL
Standard: 30 ng/mL
Detection Range: 0.47-30 ng/mL
Sample type: Serum, plasma, tissue homogenates and other biological fluids
Assay length: 2.5h
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 Caffeine. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Caffeine. 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 Caffeine 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
30.00 0.189
15.00 0.386
7.50 0.575
3.75 0.731
1.88 0.977
0.94 1.511
0.47 1.799
0.00 2.028

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 Caffeine and the recovery rates were calculated by comparing the measured value to the expected amount of Caffeine in samples.
Matrix Recovery range Average
serum(n=5) 90-105% 97%
EDTA plasma(n=5) 85-97% 91%
Heparin plasma(n=5) 96-107% 101%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Caffeine 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) 79-95% 96-105% 85-96% 90-99%
EDTA plasma(n=5) 78-92% 83-96% 95-107% 87-102%
Heparin plasma(n=5) 85-107% 84-98% 95-103% 86-98%