Multiplex Assay Kit for Cytochrome P450 27B1 (CYP27B1), etc. by FLIA (Flow Luminescence Immunoassay)
Size: 96T
Catalogue Number: LMD539Mu-96
Citations, Manuals and MSDS Available upon request.
Detection range: 0.04-40ng/mL
Application: FLIA Kit for Antigen Detection.
Organism species: Mus musculus (Mouse)
Sensitivity: The minimum detectable dose of this kit is typically less than 0.013 ng/mL
Sample type: Tissue homogenates, cell lysates, cell culture supernates and other biological fluids
Assay length: 3h
Method: Double-antibody Sandwich
Specificity: This assay has high sensitivity and excellent specificity for detection of Cytochrome P450 27B1 (CYP27B1), etc. by FLIA (Flow Luminescence Immunoassay).No significant cross-reactivity or interference between Cytochrome P450 27B1 (CYP27B1), etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.
Precision: Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Cytochrome P450 27B1 (CYP27B1), etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Cytochrome P450 27B1 (CYP27B1), etc. by FLIA (Flow Luminescence Immunoassay) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%
Assay procedure summary: 1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.
Test principle: Analyte-specific antibodies are pre-coated onto color-coded microparticles. Microparticles, standards, and samples are pipetted into wells and the immobilized antibodies bind the analytes of interest. After washing away any unbound substances, a biotinylated antibody cocktail specific to the analytes of interest is added to each well. Following a wash to remove any unbound biotinylated antibody, Streptavidin-Phycoerythrin conjugate (Streptavidin-PE), which binds to the biotinylated detection antibodies, is added to each well. A final wash removes unbound Streptavidin-PE and the microparticles are resuspended in buffer and read using the Luminex or Bio-Plex analyzer.The MFI developed isproportional to the concentration of analytes of interest in the sample.
Alternative Names: VDD1, PDDR; CYP1, P450c1; 25-Hydroxyvitamin D3 1-Alpha-Hydroxylase; Cytochrome P450 Family 27 Subfamily B Polypeptide 1; Calcidiol 1-monooxygenase; VD3 1A hydroxylase
Item Name: Cytochrome P450 27B1
Research Field: Metabolic pathway;
Uniprot: O35084
Research Use Only