BCIP/NBT Chromogen-Substrate, Ready-to-use, 100mls
Alkaline Phosphatase based immunostaining systems are gaining wide acceptance among pathologists because of their relatively high signal to noise ratio. 5-bromo,4-chloro,3-indolylphosphate (BCIP)/nitroblue tetrazolium (NBT) substrate for alkaline phosphatase-based detection systems, is a commonly used substrate chromogen. BCIP acts as the substrate for alkaline phosphatase, and the NBT enhances the purplish-brown final color of the precipitates at the reaction sites. This chromogen is useful for immunohistochemistry, immunocyto-chemistry blots and in situ hybridization procedures in both manual and automated systems.
BCIP/NBT is compatible with organic solvents so that it can be counterstained with Nuclear Fast Red or Methylene Green. The slides can be dehydrated and mounted with Clariøn (Cat. No. M05, M06, M07) or other organic solvent-based mounting medium.
For better contrast counterstaining, the BC-50 Red counterstain (Cat. No. M11) can be used. Slides should be mounted in Situ/Mount (Cat. No. M04) when BC-50 Red is used as this counterstain is not compatible with other mounting media.
Refrigerate at 4°C. Do not freeze.
Using the Chromogen-Substrate:
Remove the BCIP/NBT Chromogen-Substrate from the refrigerator 20 minutes before application.
1. Apply Ready-to-Use BCIP/NBT Chromogen Substrate to the specimen. Before applying this solution make sure to wash the specimens extensively with buffer to remove tracing reagent containing alkaline phosphatase.
2. Incubate for 15-30 minutes at room temperature. Wash specimens with distilled water and proceed with the counterstaining steps.