Principle of the Assay
The microtiter plate provided in this kit has been pre-coated with an antibody specific to AOX1. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific to AOX1. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After the TMB substrate solution is added, only those wells that contain AOX1, biotin-conjugated antibody, and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution, and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of AOX1 in the samples is then determined by comparing the O.D. of the samples to the standard curve.
For Use with serum, plasma, and cell culture supernatants. For Research Use Only. Not for use in diagnostic procedures.
Oxidase with broad substrate specificity, oxidizing aromatic azaheterocycles, such as N1-methylnicotinamide, N-methylphthalazinium and phthalazine, as well as aldehydes, such as benzaldehyde, retinal, pyridoxal, and vanillin. Plays a key role in the metabolism of xenobiotics and drugs containing aromatic azaheterocyclic substituents. Is probably involved in the regulation of reactive oxygen species homeostasis. May be a prominent source of superoxide generation via the one-electron reduction of molecular oxygen. May also catalyze nitric oxide (NO) production via the reduction of nitrite to NO with NADH or aldehyde as electron donor. May play a role in adipogenesis.
AO; AOH1; Azaheterocycle hydroxylase
|Kit Components||96 Wells Quantity/Size|
|Pre-coated, ready-to-use 96-well strip plate||1 plate|
|Plate sealer for 96 wells||2|
|Diluent buffer||1 bottle|
|Detection Reagent A||1 bottle|
|Detection Reagent B||1 bottle|
|TMB Substrate||1 tube|
|Stop Solution||1 tube|
|Wash Buffer (30 ℅ concentrate)||1 tube|
|Product data sheet||1 copy|
|Storage||The TMB Substrate, Wash Buffer (30X concentrate), and the Stop Solution should be stored at 4°C upon receipt, while the other items should be stored at -20°C.|
Intra-assay Precision (Precision within an assay): 3 samples with low, middle, and high-level AOX1 were tested 20 times on one plate, respectively.
|SENSITIVITY||The minimum detectable dose was 0.107ng/mL.|
|SPECIFICITY||This assay has high sensitivity and excellent specificity for the detection of AOX1.
No significant cross-reactivity or interference between AOX1 and analogs was observed.
Limited by current skills and knowledge, it is impossible to perform all possible cross-reactivity detection tests between AOX1 and all analogs, therefore, cross reactivity may still exist.