Description:
Apoptosisisacellularprocessinvolvingcytoplasmiccondensationandcellshrinkage,chromatincondensation,membraneblebbing,nuclearfragmentation,andtheformationofapoptoticbodies,whichleadstoprogrammedcelldeath.Duringtheprocess,aseriesofbiochemicalreactionsoccur,suchastheactivationofintracellularproteasesandregulationofapoptosis-associatedgenes.Signosis’Apoptosis-AssociatedCDNAPlateArrayprovidesanapproachforprofilingtheexpressionofapoptosis-regulatinggenes.ApplicableGrid:
ListofApplicableGenes
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | |
| A | AIF | BID | GAPDH | AIF | BID | GAPDH | AIF | BID | GAPDH | AIF | BID | GAPDH |
| B | APAF1 | CASP3 | LTA | APAF1 | CASP3 | LTA | APAF1 | CASP3 | LTA | APAF1 | CASP3 | LTA |
| C | BAD | CASP8 | MCL1 | BAD | CASP8 | MCL1 | BAD | CASP8 | MCL1 | BAD | CASP8 | MCL1 |
| D | BAK | CASP9 | p53 | BAK | CASP9 | p53 | BAK | CASP9 | p53 | BAK | CASP9 | p53 |
| E | BAX | CYCS | SMAC | BAX | CYCS | SMAC | BAX | CYCS | SMAC | BAX | CYCS | SMAC |
| F | BCL2 | FAS | Survivin | BCL2 | FAS | Survivin | BCL2 | FAS | Survivin | BCL2 | FAS | Survivin |
| G | Bcl-xL | FASL | TNFa | Bcl-xL | FASL | TNFa | Bcl-xL | FASL | TNFa | Bcl-xL | FASL | TNFa |
| H | Beta-actin | FLIP | Blank | Beta-actin | FLIP | Blank | Beta-actin | FLIP | Blank | Beta-actin | FLIP | Blank |
Principle:
Signosis’proprietarycDNAplatearrayisaplate-basedhybridizationprofilinganalysisformonitoringtheexpressionofdozensofgenesthroughreversetranscriptionofmRNAintocDNA.Likearrayanalyses,totalRNAisfirstreversetranscribedintocDNAinthepresenceofbiotin-dUTPintheassay.Targetedgenesarethenspecificallycapturedontoindividualwellsonaplate,insteadofmembranes,throughapre-coatedgene-specificoligonucleotide.ThecapturedcDNAsarefurtherdetectedwithstreptavidin-HRP.Luminescenceisreportedasrelativelightunits(RLUs)onamicroplateluminometer.Theexpressionlevelofgenesisdirectlyproportionaltotheluminescentintensity. |
Data:
ApoptosiscDNAarrayanalysisofgeneexpressioninHeLa.HeLacellweretreatedwithandwithout 2uMcamptothcinfor16hours. TotalRNAswerepreparedforthecDNAplatearrayassay.
