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Bb(Bilirubin) ELISA Kit

SKU: E-EL-0076

  • $ 42095
  • Save $ 7500


Bb(Bilirubin) ELISA Kit

Detection Range 0.31-20 μg/mL
Sensitivity 0.19 μg/mL

 

Product Details

Properties

Assay Type Competitive-ELISA
size 96T 
Assay Time 2.5h
Detection Method Colormetric
Detection Range 0.31-20 μg/mL
Sensitivity 0.19 μg/mL
Sample Volume 50μL
Sample Type serum, plasma and other biological fluids
Specificity This kit recognizes Universal Bb in samples.No significant cross-reactivity or interference between Universal Bb 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 Bb 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 Bb. During the reaction, Universal Bb in the sample or standard competes with a fixed amount of Universal Bb on the solid phase supporter for sites on the Biotinylated Detection Ab specific to Universal Bb. 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 Bb 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 Bb were tested 20 times on one plate, respectively.
    Inter-assay Precision (Precision between assays): 3 samples with low, mid range and
    high level Bb 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) 4.9 11.5 45.4 5.1 10.6 48.2
    Standard deviation 0.3 0.7 2.1 0.3 0.4 2.5
    CV (%) 6.12 6.09 4.63 5.88 3.77 5.19


    Recovery

    The recovery of Bb 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) 87-103 94
    EDTA plasma(n=8) 88-102 94
    Cell culture media(n=8) 90-107 97


    Target Information

    Research Area 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