Overview | Themostsensitivealkalinephosphatasesubstrateavailableforblottingtechniquesandinsituhybridizationchemistryisthe5-bromo-4-chloro-3-indolylphosphate/p-nitrobluetetrazoliumsystem(BCIP/NBT).UsingproprietaryproductionandstABIlizationprocedures,CHEMICONhasmadeavailableauniqueBCIP/NBTreagentthatisstableatroomtemperatureandseveraltimesmoresensitivethanexistingformulations.ForlocalizationofenzymelabeledprobesfollowingWestern,Northern,Southernordotblotting,thereagentisaddeddirectlytothemembrane.Followinga10-15minuteincubation,thereactionisterminatedbydistilledwaterrinses.Longerincubationsmaybeemployedifnecessary,butuseofbiotin/streptavidin-alkalinephosphatasesignalingcomplexeswillnegateprolongedreactiontimes. |