Enzyme-linked immunosorbent assay for Antigen Detection.
Organism Species
Rattus norvegicus (Rat)
Format
48T, 96T, 96Tx5, 96Tx10, 96Tx100
Specificity
This assay has high sensitivity and excellent specificity for detection of Afamin (AFM) . No significant cross-reactivity or interference between Afamin (AFM) and analogues was observed.
Sensitivity
The minimum detectable dose of this kit is typically less than 0.93ng/mL
Detection Range
2.47-200ng/mL
Precision
Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Afamin (AFM) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Afamin (AFM) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV Inter-Assay: CV Inter-Assay: CV
Stability
The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.
Assay Principle
The microtiter plate provided in this kit has been pre-coated with an antigen. Standards or samples are then added to the appropriate microtiter plate wells with a Horseradish Peroxidase (HRP) -conjugated secondary antibody. After TMB substrate solution is added, those wells that contain Afamin (AFM) 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 Afamin (AFM) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
Assay Protocol
1. Prepare all reagents, samples and standards; 2. Add 50µ L standard or sample to each well. And then add 50µ L prepared Detection Reagent A immediately. Shake and mix. Incubate 1 hour at 37° C; 3. Aspirate and wash 5 times; 4. Add 90µ L Substrate Solution. Incubate 10-20 minutes at 37° C; 5. Add 50µ L Stop Solution. Read at 450 nm immediately.
