Description:
ThecytochromeP450familyofenzymesplaycentralrolesintheoxidativemetabolismofawiderangeofxenobiotics,includingchemicals,sterols,fattyacidsandanticancerdrugs. HumanCYPgenesencodefifty-sevenP450proteinswithnumeroushomologvariantsindifferenttissues.Theyarehighlydifferentiallyexpressedinresponsetoavarietyofenvironmentalchemicalsandpharmaceuticals.ApplicableGrid:
ListofApplicableGenes
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | |
| A | CYP1a1 | Cyp2d22 | CYP2s1 | CYP1a1 | Cyp2d22 | CYP2s1 | CYP1a1 | Cyp2d22 | CYP2s1 | CYP1a1 | Cyp2d22 | CYP2s1 |
| B | Cyp1a2 | Cyp2d26 | Cyp3a13 | Cyp1a2 | Cyp2d26 | Cyp3a13 | Cyp1a2 | Cyp2d26 | Cyp3a13 | Cyp1a2 | Cyp2d26 | Cyp3a13 |
| C | Cyp1b1 | Cyp2d40 | Cyp3a16 | Cyp1b1 | Cyp2d40 | Cyp3a16 | Cyp1b1 | Cyp2d40 | Cyp3a16 | Cyp1b1 | Cyp2d40 | Cyp3a16 |
| D | Cyp2a12 | Cyp2d9 | Cyp3a41a | Cyp2a12 | Cyp2d9 | Cyp3a41a | Cyp2a12 | Cyp2d9 | Cyp3a41a | Cyp2a12 | Cyp2d9 | Cyp3a41a |
| E | CYP2a22 | CYP2d9 | Cyp4a32 | CYP2a22 | CYP2d9 | Cyp4a32 | CYP2a22 | CYP2d9 | Cyp4a32 | CYP2a22 | CYP2d9 | Cyp4a32 |
| F | Cyp2b10 | Cyp2e1 | Cyp4b1 | Cyp2b10 | Cyp2e1 | Cyp4b1 | Cyp2b10 | Cyp2e1 | Cyp4b1 | Cyp2b10 | Cyp2e1 | Cyp4b1 |
| G | Cyp2b13 | Cyp2f2 | 18S-rRNA | Cyp2b13 | Cyp2f2 | 18S-rRNA | Cyp2b13 | Cyp2f2 | 18S-rRNA | Cyp2b13 | Cyp2f2 | 18S-rRNA |
| H | Cyp2d10 | Cyp2j6 | Blank | Cyp2d10 | Cyp2j6 | Blank | Cyp2d10 | Cyp2j6 | Blank | Cyp2d10 | Cyp2j6 | Blank |
Principle:
Signosis’proprietaryCDNAplatearrayisaplate-basedhybridizationprofilinganalysisformonitoringtheexpressionofdozensofgenesthroughreversetranscriptionofmRNAintocDNA.Likearrayanalyses,totalRNAisfirstreversetranscribedintocDNAinthepresenceofbiotin-dUTPintheassay.Targetedgenesarethenspecificallycapturedontoindividualwellsonaplate,insteadofmembranes,throughapre-coatedgene-specificoligonucleotide.ThecapturedcDNAsarefurtherdetectedwithstreptavidin-HRP.Luminescenceisreportedasrelativelightunits(RLUs)onamicroplateluminometer.Theexpressionlevelofgenesisdirectlyproportionaltotheluminescentintensity.
Data:
AnalysisofexpressionofP450geneswithP450cDNAPlateArray. 2ugofRNAsfromratbrainwassubjectedtoP450cDNAplateassay.ThecDNAplatearrayweredetectedwithHRPbyaluminometerplatereader.
