This ELISA kit can be applied to the in vitro quantitative determination of Cotinine concentrations in serum, plasma and other biological fluids.
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 Â± 2 nm. The concentration of Cotinine in the samples is then determined by comparing the OD of the samples to the standard curve.
|Detection Range||0.63â40 ng/mL|
|Sample Type||Serum, plasma and other biological fluids|
This kit recognizes Cotinine in samples. No significant cross-reactivity or interference was observed.
The unopened kit can be stored at 4â 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.
|Micro ELISA Plate(Dismountable)||8 wells Ã12 strips||-20â, 6 months|
|Reference Standard||2 vials|
|Concentrated Biotinylated Detection Ab (100Ã)||1 vial, 120 Î¼L|
|Concentrated HRP Conjugate (100Ã)||1 vial, 120 Î¼L||-20â(shading light), 6 months|
|Reference Standard & Sample Diluent||1 vial, 20 mL||4â, 6 months|
|Biotinylated Detection Ab Diluent||1 vial, 14 mL|
|HRP Conjugate Diluent||1 vial, 14 mL|
|Concentrated Wash Buffer (25Ã)||1 vial, 30 mL|
|Substrate Reagent||1 vial, 10 mL||4â(shading light)|
|Stop Solution||1 vial, 10 mL||4â|
|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.
1.Add 50 Î¼L of standard or sample to each well.
2.Immediately add 50 Î¼L of Biotinylated Detection Ab to each well.
3.Incubate for 45 min at 37â. Aspirate and wash 3 times.
4.Add 100 Î¼L of HRP Conjugate to each well. Incubate for 30 min at 37â. Aspirate and wash 5 times.
5.Add 90 Î¼L of Substrate Reagent. Incubate for 15 min at 37â.
6.Add 50 Î¼L of Stop Solution.
7.Read the OD at 450 nm immediately. Calculation of the results.