This ELISA kit can be applied to the in vitro quantitative determination of 3-NT 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 3-NT. During the reaction, 3-NT in the sample or standard competes with a fixed amount of 3-NT on the solid phase supporter for sites on the Biotinylated Detection Ab specific to 3-NT. 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 3-NT in the samples is then determined by comparing the OD of the samples to the standard curve.
|Detection Range||1.56—100 ng/mL|
|Sample Type||Serum, plasma and other biological fluids|
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.