Description:
ThecytochromeP450familyofenzymesplaycentralrolesintheoxidativemetabolismofawiderangeofxenobiotics,includingchemicals,sterols,fattyacidsandanticancerdrugs. HumanCYPgenesencodefifty-sevenP450proteinswithnumeroushomologvariantsindifferenttissues.Theyarehighlydifferentiallyexpressedinresponsetoavarietyofenvironmentalchemicalsandpharmaceuticals.ApplicableGrid:
ListofApplicableGenes
| 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’proprietaryCDNAplatearrayisaplate-basedhybridizationprofilinganalysisformonitoringtheexpressionofdozensofgenesthroughreversetranscriptionofmRNAintocDNA.Likearrayanalyses,totalRNAisfirstreversetranscribedintocDNAinthepresenceofbiotin-dUTPintheassay.Targetedgenesarethenspecificallycapturedontoindividualwellsonaplate,insteadofmembranes,throughapre-coatedgene-specificoligonucleotide.ThecapturedcDNAsarefurtherdetectedwithstreptavidin-HRP.Luminescenceisreportedasrelativelightunits(RLUs)onamicroplateluminometer.Theexpressionlevelofgenesisdirectlyproportionaltotheluminescentintensity.
Data:
AnalysisofexpressionofP450geneswithP450cDNAPlateArray.2ugofRNAsfromMCF7,HL-60,HeLaand293celllinesweresubjectedtoP450cDNAplateassay.ThecDNAplatearrayweredetectedwithHRPbyaluminometerplatereader.
