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BCIP/NBT Substrate Kit, Alkaline Phosphatase (AP), (5-bromo-4-chloro-3-indolyl phosphate/nitroblue tetrazolium) (SK-5400)



BCIP/NBT AP Substrate Kit forms an indigo, heat stable reaction product. This chromogen can be used singly or in combination with other alkaline phosphatase or peroxidase substrates for multiple label applications.


  • Greater sensitivity than conventional substrates
  • Consistent and reliable
  • Ideal for IHC/ICC/ISH and blots
  • Fluorescent
  • Heat Stable
  • Permanent mounting
  • No wait times for mixing and dissolving powders or tablets
  • Sufficient reagents to produce 200 ml of working solution
  • Excitation: 645-685 nm*
  • Emission: peaks at 823 and 855 nm*


*Biotechniques (2007) 42:756-759 and BioTechniques (2014) 57:254-256


More Information
Unit Size1 kit
ApplicationsImmunohistochemistry / Immunocytochemistry, Immunofluorescence, In situ hybridization, Blotting Applications, Elispot
Detection Enzyme(s)Alkaline Phosphatase
Substrate Reaction ProductPrecipitating Reaction Product
Chromogen ColorIndigo
Color of FluorescenceFar Red
MicroscopyBrightfield, Fluorescence, Spectral Imaging
Additional Substrate PropertiesContrast with Tissue Pigments, Fluorescent, Heat Resistant, Multiple Labeling
MountingAqueous (Hardening), Aqueous (Non-Hardening), Non-Aqueous (Permanent)
Safety TitleWARNING: Reproductive Harm –

Kit Contents:

  • 4 ml BCIP/NBT Reagent 1
  • 3 ml BCIP/NBT Reagent 2
  • 4 ml BCIP/NBT Reagent 3


Technical Information

This substrate can be used for developing blots or for in situ hybridization. At 20-30 minute development times, the sensitivity of this substrate is equivalent to the other alkaline phosphatase substrates. However, BCIP/NBT will continue to develop from several hours to overnight making it one of the most sensitive chromogenic substrates. This kit contains stock solutions in convenient dropper bottles.

SUBSTRATEsensitivity2 jpg  Substrate.CounterstainChart jpg

SubstrateCombos jpg

Applicable patents and legal notices are available at legal notices.

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