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Cotinine ELISA Kit

Cotinine ELISA Kit
Cotinine ELISA Kit
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Product Code : E-EL-0064
Brand Name : Cotinine ELISA Kit
Product Description

Intended use

This ELISA kit can be applied to the in vitro quantitative determination of  Cotinine concentrations in serum, plasma and other biological fluids.

Test principle

This ELISA kit uses Competitive-ELISA as the method. The microtiter plate provided in this kit has been pre-coated with Cotinine. During the reaction,  Cotinine in the sample or standard competes with a fixed amount of Cotinine on the solid phase supporter for sites on the Biotinylated Detection Ab specific to  Cotinine. Excess conjugate and unbound sample or standard are washed from the plate, and Avidin conjugated to Horseradish Peroxidase (HRP) are added to each microplate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by adding Stop Solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of  Cotinine in the samples is then determined by comparing the OD of the samples to the standard curve.

Assay type Competitive
Format 96T
Assay time 2.5h
Reactivity Universal
Detection Method Colormetric
Detection Range 0.63—40 ng/mL
Sensitivity 0.38 ng/mL
Sample Volume 50μL
Sample Type Serum, plasma and other biological fluids

Specificity

This kit recognizes Cotinine in samples. No significant cross-reactivity or interference was observed.

Kit components & Storage

The unopened kit can be stored at 4℃ for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions since the kit is received.

Item Specifications Storage
Micro ELISA Plate(Dismountable) 8 wells ×12 strips -20℃, 6 months
Reference Standard 2 vials
Concentrated Biotinylated Detection Ab (100×) 1 vial, 120 μL
Concentrated HRP Conjugate (100×) 1 vial, 120 μL -20℃(shading light), 6 months
Reference Standard & Sample Diluent 1 vial, 20 mL 4℃, 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 4℃(shading light)
Stop Solution 1 vial, 10 mL 4℃
Plate Sealer 5 pieces
Product Description 1 copy
Certificate of Analysis 1 copy

Note: All reagent bottle caps must be tightened to prevent evaporation and microbial pollution.
The volume of reagents in partial shipments is a little more than the volume marked on the label, please use in measuring instead of directly pouring.

Other supplies required

  • Microplate reader with 450nm wavelength filter
  • High-precision transferpettor, EP tubes and disposable pipette tips
  • 37℃ Incubator
  • Deionized or distilled water
  • Absorbent paper
  • Loading slot for Wash Buffer

Assay Procedure

1.Add 50 μL of standard or sample to each well.

2.Immediately add 50 μL of Biotinylated Detection Ab to each well.

3.Incubate for 45 min at 37℃. Aspirate and wash 3 times.

4.Add 100 μL of HRP Conjugate to each well. Incubate for 30 min at 37℃. Aspirate and wash 5 times.

5.Add 90 μL of Substrate Reagent. Incubate for 15 min at 37℃.

6.Add 50 μL of Stop Solution.

7.Read the OD at 450 nm immediately. Calculation of the results.

 


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