Characterization
Description(s):The cytochrome P450 family of enzymes play central roles in the oxidative metabolism of a wide range of xenobiotics, including chemicals, sterols, fatty acids and anticancer drugs. Human CYP genes encode fifty-seven P450 proteins with numerous homolog variants in different tissues. They are highly differentially expressed in response to a variety of environmental chemicals and pharmaceuticals.
Signosis has developed a plate-based array for profiling 20+ P450-regulated genes. By using this assay, researchers are able to compare gene expression in up to three samples on a single microtiter plate.
Applicable Grid:
List of Applicable Genes
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | |
| A | CYP1A1 | CYP2E1 | CYP11B1 | CYP1A1 | CYP2E1 | CYP11B1 | CYP1A1 | CYP2E1 | CYP11B1 | CYP1A1 | CYP2E1 | CYP11B1 |
| B | CYP1A2 | CYP3A4 | CYP17A1 | CYP1A2 | CYP3A4 | CYP17A1 | CYP1A2 | CYP3A4 | CYP17A1 | CYP1A2 | CYP3A4 | CYP17A1 |
| C | CYP1B1 | CYP3A5/A7 | CYP24A1 | CYP1B1 | CYP3A5/A7 | CYP24A1 | CYP1B1 | CYP3A5/A7 | CYP24A1 | CYP1B1 | CYP3A5/A7 | CYP24A1 |
| D | Beta-actin | GAPDH | CYP26A1 | Beta-actin | GAPDH | CYP26A1 | Beta-actin | GAPDH | CYP26A1 | Beta-actin | GAPDH | CYP26A1 |
| E | CYP2A6 | CYP4A11 | CYP26B1 | CYP2A6 | CYP4A11 | CYP26B1 | CYP2A6 | CYP4A11 | CYP26B1 | CYP2A6 | CYP4A11 | CYP26B1 |
| F | CYP2B6 | CYP4B1 | CYP27A1 | CYP2B6 | CYP4B1 | CYP27A1 | CYP2B6 | CYP4B1 | CYP27A1 | CYP2B6 | CYP4B1 | CYP27A1 |
| G | CYP2C19 | CYP4F2/F3/F8 | CYP39A1 | CYP2C19 | CYP4F2/F3/F8 | CYP39A1 | CYP2C19 | CYP4F2/F3/F8 | CYP39A1 | CYP2C19 | CYP4F2/F3/F8 | CYP39A1 |
| H | CYP2D6 | CYP11A1 | CYP46A1 | CYP2D6 | CYP11A1 | CYP46A1 | CYP2D6 | CYP11A1 | CYP46A1 | CYP2D6 | CYP11A1 | CYP46A1 |
Principle
Signosis’ proprietary cDNA plate array is a plate-based hybridization profiling analysis for monitoring the expression of dozens of genes through reverse transcription of mRNA into cDNA. Like array analyses, total RNA is first reverse transcribed into cDNA in the presence of biotin-dUTP in the assay. Targeted genes are then specifically captured onto individual wells on a plate, instead of membranes, through a pre-coated gene-specific oligonucleotide. The captured cDNAs are further detected with streptavidin-HRP. Luminescence is reported as relative light units (RLUs) on a microplate luminometer. The expression level of genes is directly proportional to the luminescent intensity.
Data
Analysis of expression of P450 genes with P450 cDNA Plate Array. 2ug of RNAs from MCF7, HL-60, HeLa and 293 cell lines were subjected to P450 cDNA plate assay. The cDNA plate array were detected with HRP by a luminometer plate reader.
