This ELISA kit can be applied to the in vitro quantitative determination of Pregnenolone 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 Pregnenolone. During the reaction, Pregnenolone in the sample or standard competes with a fixed amount of Pregnenolone on the solid phase supporter for sites on the Biotinylated Detection Ab specific to Pregnenolone. 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 Pregnenolone in the samples is then determined by comparing the OD of the samples to the standard curve.
|Detection Range||0.78—50 ng/mL|
|Sample Type||Serum, plasma and other biological fluids|
This kit recognizes Pregnenolone in samples. No significant cross-reactivity or interference was observed.
The recovery of Pregnenolone spiked to three different levels in samples throughout the range of the assay in various matrices was evaluated.
|Sample Type||Range (%)||Average Recovery (%)|
|EDTA plasma (n=5)||86-99||93|
|Cell culture media (n=5)||85-95||90|
Samples were spiked with high concentrations of Human ACTH and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
|Serum (n=5)||EDTA plasma (n=5)||Cell culture media (n=5)|
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.
|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.