This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human ADP/Acrp30 . Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human ADP/Acrp30 and Avidin-Horseradish Peroxidase (HRP) conjugate is added to each micro plate well successively and then incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human ADP/Acrp30 , biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color turns to yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human ADP/Acrp30 . You can calculate the concentration of Human ADP/Acrp30 in the samples by comparing the OD of the samples to the standard curve.
|Assay type||Sandwich ELISA|
|Sample Volume Required Per Well||100Î¼L|
|Sample Type||serum, plasma and other biological fluids|
|Micro ELISA Plate||8 wells Ã12 strips||4â/-20â#|
|Reference Standard||2 vials||4â/-20â#|
|Reference Standard & Sample Diluent||1vial 20mL||4â|
|Concentrated Biotinylated Detection Ab||1vial 120Î¼L||4â/-20â#|
|Biotinylated Detection Ab Diluent||1vial 10mL||4â|
|Concentrated HRP Conjugate||1vial 120Î¼L||4â(shading light)|
|HRP Conjugate Diluent||1vial 10mL||4â|
|Concentrated Wash Buffer (25Ã)||1vial 30mL||4â|
|Substrate Reagent||1vial 10mL||4â(shading light)|
|Stop Solution||1vial 10mL||4â|
|Certificate of Analysis||1 copy|
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Human ADP/Acrp30 were tested on 3 different plates, 20 replicates in each plate.
|Intra-assay Precision||Inter-assay Precision|
|C V (%)||5.800||5.600||4.500||6.900||5.000||3.600|
The recovery of Human ADP/Acrp30 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)||91-103||98.000|
|Cell culture media (n=5)||94-109||100.000|
This kit recognizes recombinant and natural Human ADP/Acrp30 . No significant cross-reactivity or interference was observed.