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Specifications:
| Application | Western Blot, Colorimetric Assay | ||
| Storage Temperature | -15°C to -25°C | ||
| Product Type | Molecular Biology Reagent | Forms | Solid |
| Product Brand | Thermo Fisher Scientific™, Invitrogen | ||
| Product Grade | Molecular Biology | Formula | C₄₀H₃₀Cl₂N₁₀O₆ / C₈H₄BrClNNa₂O₄P |
The Invitrogen™ NBT/BCIP Reagent Kit provides chromogenic substrates for colorimetric detection of alkaline phosphatase (AP) activity in various applications including blotting, in situ hybridization (ISH), and immunohistochemistry (IHC). This ready-to-use kit contains 1 gram each of Nitro Blue Tetrazolium chloride (NBT) and 5-Bromo-4-chloro-3-indolyl phosphate (BCIP)—the widely used substrate pair that produces an intense blue-purple precipitate upon reaction with alkaline phosphatase.
When AP cleaves BCIP, it generates an intermediate that reacts with NBT to form a water-insoluble formazan product. This enables clear, chromogenic signal development suitable for both membrane and tissue applications, without the need for fluorescence or radioactive labeling.
Key Features and Benefits
- Ready-to-use Substrate Kit: Includes both BCIP and NBT for reliable, reproducible phosphatase detection
- Intense Color Development: Forms a blue-purple precipitate at AP-positive sites
- Ideal for Non-Fluorescent Detection: Enables colorimetric visualization of AP activity
- Versatile Application: Suitable for use in blotting membranes, tissue sections, and ISH
- Stable Storage Format: Long-term storage at –5°C to –30°C; light-protected formulation
Applications
- Western blot detection using AP-conjugated secondary antibodies
- In situ hybridization (ISH) for gene expression and localization studies
- Immunohistochemistry (IHC) in formalin-fixed or paraffin-embedded tissues
- Membrane-based assays (e.g., dot blots, Southern and Northern blots)
- Enzyme-based assays requiring visible substrate signal development
Technical Specifications
| Property | Details |
|---|---|
| Kit Contents | 1 g Nitro Blue Tetrazolium (NBT) 1 g BCIP |
| Detection Method | Colorimetric |
| Substrate Target | Alkaline Phosphatase (AP) |
| Substrate Properties | Chemical chromogenic |
| Reaction Product Color | Blue-purple precipitate |
| Storage Temperature | –5°C to –30°C |
| Shipping Conditions | Room temperature |
| Light Sensitivity | Yes – protect from light |
| Unit Size | Each kit |
The Invitrogen™ NBT/BCIP Reagent Kit is a trusted and convenient solution for researchers requiring reliable chromogenic detection of alkaline phosphatase activity in blotting and histological assays. With its intense color output and versatility across a range of molecular biology and pathology techniques, this kit provides clear, stable, and publication-quality results without the complexity of fluorescence or radioactivity.