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5-HIAA(5-Hydroxyindoleacetic Acid) ELISA Kit
ELK7838
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48 T $265.00
96 T $376.00
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Overview

Product name: 5-HIAA(5-Hydroxyindoleacetic Acid) ELISA Kit
Reactivity: General
Alternative Names: 5HIAA
Assay Type: Competitive Inhibition
Sensitivity: 1.93 ng/mL
Standard: 400 ng/mL
Detection Range: 6.25-400 ng/mL
Sample Type: serum, plasma, urine, cerebrospinal fluid and other biological fluids
Assay Length: 2h
Research Area: Metabolic pathway;Tumor immunity;Endocrinology;Urology;
Test principle: This assay employs the competitive inhibition enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with 5-HIAA protein. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to 5-HIAA. 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-HIAA 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
400.00 0.199
200.00 0.352
100.00 0.577
50.00 0.895
25.00 1.139
12.50 1.578
6.25 1.789
0.00 2.115

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-HIAA and the recovery rates were calculated by comparing the measured value to the expected amount of 5-HIAA in samples.
Matrix Recovery range Average
serum(n=5) 90-105% 97%
EDTA plasma(n=5) 85-97% 91%
Heparin plasma(n=5) 92-105% 93%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of 5-HIAA 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) 89-97% 85-94% 98-105% 92-101%
EDTA plasma(n=5) 92-101% 87-98% 85-92% 79-96%
Heparin plasma(n=5) 89-102% 88-97% 88-104% 97-108%