DIGLuminescentDetectionKit地高辛发光检测试剂盒,CSPD化学发光法检测,灵敏度可达0.03pg同源DNA;可检测50张
Application
ProductDescription
Sensitivity:0.03pgofhomologousDNAor0.1pgofhomologousRNAisdetectedinaSouthernordotblotafter<30-minexposuretime.Single-copygenesaredetectedin0.3µgmammaliangenomicDNA.
Figure1:DetectionofDIG-labelednucleicacidswithCSPD.HumangenomicDNAwasdigestedwithEcoRI,separatedona1agarosegel,andblottedontopositivelychargednylonmembranes.Theblotswerehybridizedwith50ng/mlDIG-labeledβ-actinRNA.ChemiluminescentdetectionwasperformedaccordingtothestandardDIGchemiluminescent-detectionprocedure,usingCSPData0.25-mMfinalconcentration.
BackgroundInformation
Contents
- DIG-labeledControlDNA,50µl, [5µg/ml]
- DNADilutionBuffer,1ml
- Anti-digoxigenin-AP,Fabfragments,100µl,[750U/ml]
- BlockingReagent,2x50g
- CSPD,1ml,[11.6mg/ml]
Principle
- DIG-labelednucleic-acidprobesareusedforhybridizationinvariousblotapplications.
- Afterhybridization,theblotsaresubmittedtoimmunologicaldetectionusingahighlyspecificanti-digoxigenin-antibody,conjugatedtoalkalinephosphatase.
- TheadditionofthechemiluminescencesubstrateCSPDleadstoadistinctluminescentsignalthatisrecordedonX-rayorPolaroidfilm.
- Exposuretimevariesbetween5and20min,andbecauseofthelong-lastingglow,multipleexposurescanbetakenoverseveraldays.
- BlotsdevelopedwithCSPDcanbeeasilystrippedandreprobed.
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