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5-HIAA / 5-Hydroxyindoleacetic Acid ELISA Kit - Information

The 5-HIAA / 5-Hydroxyindoleacetic Acid ELISA Kit can assay for 5-HIAA / 5-Hydroxyindoleacetic Acid in the following samples: serum, blood, plasma, cell culture supernatant and other related supernatants and tissues.

How our 5-HIAA / 5-Hydroxyindoleacetic Acid ELISA Kits Work?

The ELISA Genie ELISA (enzyme-linked immunosorbent assays) assay kits are designed for the quantitative measurement of analytes in a wide variety of samples. As today's scientists demand high quality consistent data for high impact journals, We have developed a range of sensitive, fast and reliable ELISA kit assays to meet and exceed those demands. This ELISA kit uses Competitive-ELISA as the method.

The microtiter plate provided in this kit has been pre-coated with 5-HIAA / 5-Hydroxyindoleacetic Acid. During the reaction, 5-HIAA / 5-Hydroxyindoleacetic Acid in the sample or standard competes with a fixed amount of 5-HIAA / 5-Hydroxyindoleacetic Acid on the solid phase supporter for sites on the Biotinylated Detection Antibody specific to 5-HIAA / 5-Hydroxyindoleacetic Acid. Excess conjugate and unbound sample or standard are washed from the plate, and HRP-Streptavidin(SABC) is added to each microplate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm. The concentration of 5-HIAA / 5-Hydroxyindoleacetic Acid in the samples is then determined by comparing the O.D. of the samples to the standard curve.

5-HIAA / 5-Hydroxyindoleacetic Acid ELISA Kit - Data

Product name 5-HIAA(5-Hydroxyindoleacetic Acid) ELISA Kit
Catalogue No. EU2582
Alias 5-HIAA(5-Hydroxyindoleacetic Acid)
Detection method Competitive ELISA Coated with Antigen
Application This immunoassay kit allows for the in vitro quantitative determination of 5-HIAA concentrations in serum plasma and other biological fluids.
Size 96T
Range 1.563-100ng/ml
Sensitivity < 0.938ng/ml
Storage 4'C for 6 months
Recovery Matrices listed below were spiked with certain level of 5-HIAA and the recovery rates were calculated by comparing the measured value to the expected amount of 5-HIAA in samples.
MatrixRecovery range(%)Average(%)
serum(n=5) 95-101 97
EDTA plasma(n=5) 89-105 97
UFH plasma(n=5) 85-105 97
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.
serum(n=5) 86-105% 90-101% 86-105% 88-103%
EDTA plasma(n=5) 82-98% 84-100% 82-100% 87-100%
UFH plasma(n=5) 83-95% 83-98% 80-97% 87-96%
CV(%) Intra-Assay: CV<8%Inter-Assay: CV<10%
Note For Research Use Only


ELISA Kit Technical ManualELISA Kit Technical ManualMSDS


Competitive ELISA Protocol

The below protocol is a sample protocol for a competitive ELISA kits. Competitive ELISA kits allow for the detection and quantification of an analyte in a sample. This allows the researcher to calculate the amount of analyte present in their sample. This can be very useful when looking for increases in protein concentration, phosphorylation of proteins or decreases on protein concentrations.Before adding to wells, equilibrate the SABC working solution and TMB substrate for at least 30 min at 37 °C. When diluting samples and reagents, they must be mixed completely and evenly. It is recommended to plot a standard curve for each test.

Competitive ELISA Protocol

ELISA Kit Technical Manual


1. Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells!
2. Add Sample and Biotin-detection antibody: Add 50µL of Standard, Blank, or Sample per well. The blank well is added with Sample / Standard dilution buffer. Immediately add 50 µL of Biotin-detection antibody working solution to each well. Cover with the Plate sealer we provided. Gently tap the plate to ensure thorough mixing. Incubate for 45minutes at 37°C. (Solutions are added to the bottom of micro ELISA plate well, avoid inside wall touching and foaming to the best of your ability.)
3. Wash: Aspirate each well and wash, repeating the process three times Wash by filling each well with Wash Buffer (approximately 350µL) using a squirt bottle, multi-channel pipette, manifold dispenser or automated washer. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and pat it against thick clean absorbent paper.
4. HRP-Streptavidin Conjugate(SABC): Add 100µL of SABC working solution to each well. Cover with a new Plate sealer. Incubate for 30minutes at 37°C.
5. Wash: Repeat the aspiration/wash process for five times.
6. TMB Substrate: Add 90µL of TMB Substrate to each well. Cover with a new Plate sealer. Incubate for about 15-20 minutes at 37°C. Protect from light. The reaction time can be shortened or extended according to the actual color change, but not more than 30minutes. When apparent gradient appeared in standard wells, you can terminate the reaction.
7. Stop: Add 50µL of Stop Solution to each well. Color turn to yellow immediately. The adding order of stop solution should be as the same as the substrate solution.
8. OD Measurement: Determine the optical density (OD Value) of each well at once, using a microplate reader set to 450 nm. You should open the microplate reader ahead, preheat the instrument, and set the testing parameters.
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