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5-HIAA(5-Hydroxyindoleacetic Acid) ELISA Kit– MSE Supplies LLC

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5-HIAA(5-Hydroxyindoleacetic Acid) ELISA Kit

SKU: E-EL-0075

  • $ 42095


5-HIAA(5-Hydroxyindoleacetic Acid) ELISA Kit

Detection Range 1.56-100 ng/mL
Sensitivity 0.94 ng/mL

 

Product Details

Properties

Assay Type Competitive-ELISA
size 96T 
Assay Time 2.5h
Detection Method Colormetric
Detection Range 1.56-100 ng/mL
Sensitivity 0.94 ng/mL
Sample Volume 50μL
Sample Type serum, plasma and other biological fluids
Specificity This kit recognizes Universal 5-HIAA in samples.No significant cross-reactivity or interference between Universal 5-HIAA and analogues was observed
Precision Both intra-CV and inter-CV are < 10%.
Recovery 80%-120%
Introduction This ELISA kit applies to the in vitro quantitative determination of 5-HIAA concentrations in serum, plasma and other biological fluids.
Applications ELISA
Storage 2-8℃/-20℃


Test Principle

This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with Universal 5-HIAA. During the reaction, Universal 5-HIAA in the sample or standard competes with a fixed amount of Universal 5-HIAA on the solid phase supporter for sites on the Biotinylated Detection Ab specific to Universal 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 Universal 5-HIAA in tested samples can be calculated by comparing the OD of the samples to the standard curve.

Kit Components and 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 items separately according to the following conditions once the kit is received.

Item Specifications Storage
Micro ELISA Plate(Dismountable) 96T: 8 wells ×12 strips -20℃, 6 months
48T: 8 wells ×6 strips
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 -20℃(Protect from
light), 6 months
48T: 1 vial, 60 μL
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,
    respectively.

    Intra-assay Precision Inter-assay Precision
    Sample 1 2 3 1 2 3
    n 20 20 20 20 20 20
    Mean (ng/mL) 204.2 588.5 1720.2 221.1 588.2 1852
    Standard deviation 10.6 29.4 61.9 13.9 24.7 59.3
    CV (%) 5.19 5 3.6 6.29 4.2 3.2


    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


    Target Information

    Research Area Neuroscience, Metabolism

     

    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