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5-HIAA(5-Hydroxyindoleacetic Acid) ELISA Kit

Cat.No.:E-EL-0075
Reactivity: Universal

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96T
48T
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96T*5
96T*10

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Product Details

Properties

Assay type	Competitive-ELISA
Format	96T/48T
Assay time	2.0h
Detection range	1.56-100 ng/mL
Sensitivity	0.94 ng/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 50μL
Specificity	This kit recognizes 5-HIAA in samples. No significant cross-reactivity or interference between 5-HIAA and analogues was observed.
Reproducibility	Both intra-CV and inter-CV are < 10%.
Application	This ELISA kit applies to the in vitro quantitative determination of 5-HIAA concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with 5-HIAA. During the reaction, 5-HIAA in the sample or standard competes with a fixed amount of 5-HIAA on the solid phase supporter for sites on the Biotinylated Detection Ab specific to 5-HIAA. Excess conjugate and unbound sample or standard are washed away, and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns from blue to yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of 5-HIAA in tested samples can be calculated by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.

Item	Specifications	Storage
Micro ELISA Plate(Dismountable)	96T: 8 wells ×12 strips 48T: 8 wells ×6 strips	-20℃, 6 months
Reference Standard	96T: 2 vials 48T: 1 vial
Concentrated Biotinylated Detection Ab (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL	-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°C, 6 months
Biotinylated Detection Ab Diluent	1 vial, 14 mL
HRP Conjugate Diluent	1 vial, 14 mL
Concentrated Wash Buffer (25×)	1 vial, 30 mL
Substrate Reagent	1 vial, 10 mL	2-8℃(Protect from light)
Stop Solution	1 vial, 10 mL	2-8℃
Plate Sealer	5 pieces
Manual	1 copy
Certificate of Analysis	1 copy

Technical Data

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level 5-HIAA were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level 5-HIAA were tested on 3 different plates, 20 replicates in each plate.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (ng/mL)	204.20	588.50	1720.20	221.10	588.20	1852.00
Standard deviation	10.60	29.40	61.90	13.90	24.70	59.30
CV (%)	5.19	5.00	3.60	6.29	4.20	3.20

Recovery

The recovery of 5-HIAA 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)	85-98	91
EDTA plasma(n=8)	93-106	98
Cell culture media(n=8)	92-106	99

Linearity

Samples were spiked with high concentrations of 5-HIAA and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Research Area

Neuroscience, Metabolism

Assay Procedures

	1. Add 50μL standard or sample to the wells, immediately add 50μL Biotinylated Detection Ab working solution to each well. Incubate for 45 min at 37°C
	2. Aspirate and wash the plate for 3 times
	3. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	4. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	5. Add 50μL Stop Solution
	6. Read the plate at 450nm immediately. Calculation of the results

Citations

Food & Function (2021) IF: 5.396
Neuroprotective effects of macamide from maca (Lepidium meyenii Walp.) on corticosterone-induced hippocampal impairments through anti-inflammatory, neurotrophic, and synaptic protection properties

DOI: 10.1039/D1FO01720A

Sample: hippocampu
Foods (2023) IF: 5.561
Fermented Wheat Germ Alleviates Depression-like Behavior in Rats with Chronic and Unpredictable Mild Stress

DOI: 10.3390/foods12050920

Sample: hippocampus
PLoS One (2016) IF: 3.057
M30 Antagonizes Indoleamine 2,3-Dioxygenase Activation and Neurodegeneration Induced by Corticosterone in the Hippocampus

DOI: 10.1371/journal.pone.0166966

Sample: Tissue homogenate
Brain Sciences (2020) IF: 3.332
Antidepressant-Like Properties of Fish Oil on Postpartum Depression-Like Rats Model: Involvement of Serotonergic System

DOI: 10.3390/brainsci10100733

PMID: 33066310
Sample: Oil
JOURNAL OF ETHNOPHARMACOLOGY (2019) IF: 3.414
Alkaloids from the rhizomes of Ligusticum striatum exert antimigraine effects through regulating 5-HT1B receptor and c-Jun

DOI: 10.1016/j.jep.2019.03.026

PMID: 30880256
Sample: Tissue homogenate
PHARMACOLOGY BIOCHEMISTRY AND BEHAVIOR (2015) IF: 2.781
Antidepressant-like effects of the xanthine oxidase enzyme inhibitor allopurinol in rats. A comparison with fluoxetine

DOI: 10.1016/j.pbb.2015.09.016

Sample: Plasma
PLoS One (2017) IF: 2.806
Monoamine oxidase A upregulated by chronic intermittent hypoxia activates indoleamine 2,3-dioxygenase and neurodegeneration

DOI: 10.1371/journal.pone.0177940

Sample: Tissue homogenate

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