Further Information
CP27; CTX; CYP27; Sterol 27-Hydroxylase,Cerebrotendinous Xanthomatosis; Vitamin D(3) 25-hydroxylase; 5-beta-cholestane-3-alpha,7-alpha,12-alpha-triol 27-hydroxylase
3h
1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 2 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.
0.156-10ng/mL
48T, 96T, 96T?5, 96T?10, 96T?100
Cytochrome P450 27A1
Double-antibody Sandwich
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Cytochrome P450 27A1 (CYP27A1) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Cytochrome P450 27A1 (CYP27A1) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%
Metabolic pathway;
Tissue homogenates, cell lysates and other biological fluids.
The minimum detectable dose of this kit is typically less than 0.059ng/mL
This assay has high sensitivity and excellent specificity for detection of Cytochrome P450 27A1 (CYP27A1).
No significant cross-reactivity or interference between Cytochrome P450 27A1 (CYP27A1) and analogues was observed.
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.
The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Cytochrome P450 27A1 (CYP27A1). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Cytochrome P450 27A1 (CYP27A1). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Cytochrome P450 27A1 (CYP27A1), 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 Cytochrome P450 27A1 (CYP27A1) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
Q02318