Catalog No: RE10166
5-HT(5-Hydroxytryptamine) ELISA Kit
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48T

$299.00

96T

$399.00

Product Details

Species:

Universal

Alternative Name:

5HT; Serotonin; N-Methyl-Gamma

Assay Type:

Competitive

unit:

ng/mL

Standard:

900 ng/mL

Sensitivity:

3.97 ng/mL

Detection Range:

14.07-900 ng/mL

Assay Time:

1.5H

Sample Type:

Serum, Plasma, Tissue homogenates

Research:

Signal transduction;Infection immunity;Neuro science;Gastroenterology;

Test Principle:

This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with 5-HT. Standards or samples are added to the appropriate micro ELISA plate wells then with a biotin-conjugated antibody specific to 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 5-HT in the samples is then determined by comparing the OD of the samples to the standard curve.

Technical Data

As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
ng/mL OD Corrected
900 0.423
450 0.539
225 0.735
112.5 1.02
56.25 1.358
28.13 1.673
14.06 1.909
0 2.428

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level were tested on 3 different plates, 20 replicates in each plate, respectively.

Intra-assay Precision Inter-assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean(ng/mL) 17.72 37.82 252.81 18.44 28.3 314.69
Standard deviation 1.89 4.17 13.06 2.35 5.19 16.25
C V (%) 6.14 6.24 5.65 6.82 5.34 4.2

Rate of recovery

The recovery of spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
Sample Type Range(%) Average Recovery(%)
Serum (n=8) 87-95 91
EDTA plasma (n=8) 81-95 88
Cell culture media (n=8) 83-97 90

Linear

Samples were spiked with high concentrations of target proteins and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
Serum (n=5) EDTA plasma (n=5) Cell culture media (n=5)
1:2 Range (%) 85-94 85-103 87-98
Average (%) 87 101 91
1:4 Range (%) 90-99 85-103 80-92
Average (%) 92 87 85
1:8 Range (%) 82-101 83-96 96-105
Average (%) 82 94 104
1:16 Range (%) 91-102 95-102 86-97
Average (%) 96 97 92

Assay Procedures

Description