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5-HIAA(5-Hydroxyindoleacetic Acid) ELISA Kit

  • Cat.No.:E-EL-0075

  • Reactivity: Universal

To Purchase E-EL-0075

Size:
  • 96T
  • 48T
  • 24T
  • 96T*5
  • 96T*10
Price: $495
Qty:

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.
ItemSpecificationsStorage
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

Synonyms2-(5-Hydroxy-1H-indol-3-yl)acetic acid

Assay Procedures

elisa assay procedure 1

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

elisa assay procedure 2

2. Aspirate and wash the plate for 3 times

elisa assay procedure 3

3. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times

elisa assay procedure 4

4. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C

elisa assay procedure 5

5. Add 50μL Stop Solution

elisa assay procedure 6

6. Read the plate at 450nm immediately. Calculation of the results

Related Products

Elabscience® provides ancillary reagents required for ELISA kit developments.11 reagents in total, including reagents for coating, blocking, washing, improving the stability and reducing matrix interference etc.
Cat.No. Product Name Specifications
E-ELIR-001 ELISA Plate Coating Buffer(1×) 100mL, 1000mL
E-ELIR-002 ELISA Plate Coating Buffer(5×) 100mL, 1000mL
E-ELIR-003 ELISA Plate Blocking Buffer 100mL, 1000mL
E-ELIR-004 Wash Buffer for Sandwich-ELISA(25×) 100mL, 1000mL
E-ELIR-005 Wash Buffer for Competitive-ELISA(25×) 100mL, 1000mL
E-ELIR-006 Stop Solution 100mL, 1000mL
E-ELIR-007 HRP-conjugate Stabilizer 10mL, 100mL, 500mL
E-ELIR-008 HRP-conjugate Diluent 100mL, 1000mL
E-ELIR-009 Biotinylated Antibody Stabilizer 10mL, 100mL, 500mL
E-ELIR-0010 Biotinylated Antibody Diluent 10mL, 100mL, 500mL
E-ELIR-0011 Sample Diluent 10mL, 100mL, 500mL

Citations

From now on, if you have published a paper by using any of our products since 1/1/2019, fill out the “Elabscience Publication Reward Application Form”carefully and send it to orders@elabscience.com, we will get back to you with the reward after we confirm it ASAP!

View more details about our Publication Reward >>

  1. Journal of ethnopharmacology (2019) IF: 3.115
    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
  2. 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
  3. 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

    PMID: 28599322

    Sample: Tissue homogenate
  4. Plos One (2016) IF: 2.806
    M30 Antagonizes Indoleamine 2,3-Dioxygenase Activation And Neurodegeneration Induced By Corticosterone In The Hippocampus.

    DOI: 10.1371/journal.pone.0166966

    PMID: 27870896

    Sample: Tissue homogenate
  5. Pharmacology Biochemistry and Behavior (2015) IF: 2.748
    Antidepressant-Like Effects Of The Xanthine Oxidase Enzyme Inhibitor Allopurinol In Rats. A Comparison With Fluoxetine.

    DOI: 10.1016/j.pbb.2015.09.016

    PMID: 26409178

    Sample: Plasma
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