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ELK Biotechnology

ELK9416

Zebrafish 5-HT(5-Hydroxytryptamine) ELISA Kit

Zebrafish 5-HT(5-Hydroxytryptamine) ELISA Kit

Sensitivity

Zebrafish 5-HT(5-Hydroxytryptamine) ELISA Kit

Overview

Product name: Zebrafish 5-HT(5-Hydroxytryptamine) ELISA Kit
Reactivity: Zebrafish
Alternative Names: 5HT; Serotonin; N-Methyl-Gamma
Assay Type: Competitive Inhibition
Sensitivity: 3.97 ng/mL
Standard: 900 ng/mL
Detection Range: 14.07-900 ng/mL
Sample Type: serum, plasma, tissue homogenates
Assay Length: 2h
Research Area: Signal transduction;Infection immunity;Neuro science;Gastroenterology;
Test principle: This assay employs the competitive inhibition enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with Zebrafish 5-HT protein. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Zebrafish 5-HT. 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 Zebrafish 5-HT 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
900.00 0.232 —
450.00 0.385 —
225.00 0.696 —
112.50 0.823 —
56.25 1.275 —
28.13 1.592 —
14.07 1.887 —
0.00 2.123 —

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 5-HT and the recovery rates were calculated by comparing the measured value to the expected amount of 5-HT in samples.
Matrix Recovery range Average
serum(n=5) 85-99% 92%
EDTA plasma(n=5) 90-105% 97%
Heparin plasma(n=5) 95-107% 101%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of 5-HT 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) 89-102% 87-95% 85-96% 92-102%
EDTA plasma(n=5) 81-92% 86-93% 91-103% 81-94%
Heparin plasma(n=5) 86-98% 92-101% 95-104% 87-96%
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