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This ELISA kit can be applied to the in vitro quantitative determination of Human ACTH 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 Human ACTH. During the reaction, Human ACTH in the sample or standard competes with a fixed amount of Human ACTH on the solid phase supporter for sites on the Biotinylated Detection Ab specific to Human ACTH. 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 Human ACTH in the samples is then determined by comparing the OD of the samples to the standard curve.
|Detection Range||15.63â1000 pg/mL|
|Sample Type||Serum, plasma and other biological fluids|
This kit recognizes Human ACTH in samples. No significant cross-reactivity or interference was observed.
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Human ACTH were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Human ACTH were tested on 3 different plates, 20 replicates in each plate.
|Intra-assay Precision||Inter-assay Precision|
|C V (%)||7.00||5.19||3.10||5.10||5.13||4.39|
The recovery of Human ACTH 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)||85-100||91|
|Cell culture media (n=5)||86-98||93|
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.