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| Product Name | Alkaline Phosphatase Stain Kit |
| Catalog No. | RESK-0146 |
| Description | Alkaline phosphatase (AP/ALP/AKP) is a group of hydrolase isoenzymes that remove phosphate groups from substrate molecules by hydrolyzing phosphomonoesters, generating phosphate ions and free hydroxyl groups; under alkaline conditions, nucleotides, proteins and alkaloids serve as dephosphorylation substrates. Alkaline phosphatases are widely used, for example, for antibody labeling in protein and nucleic acid detection and for dephosphorylation of DNA or RNA 5' and 3' ends. This kit provides a BCIP/NBT-based chromogenic system for sensitive colorimetric detection of alkaline phosphatase activity in cells and tissues, and can also be used for chromogenic detection of ALP-conjugated membranes in Western blotting. |
| Intended Use | Colorimetric staining of alkaline phosphatase activity in cells or tissues, including identification of induced pluripotent stem cells (iPSCs), staining of osteoblasts after osteogenic differentiation, detection of endogenous alkaline phosphatase in cells or tissues, and chromogenic detection of alkaline phosphatase-conjugated membranes in Western blotting. Not intended for cell viability assays. For research use only. |
| Principle / Technology | BCIP/NBT is a commonly used substrate pair for alkaline phosphatase. Under the action of alkaline phosphatase, BCIP is hydrolyzed, and the hydrolysis product reacts with NBT to form an insoluble dark blue to blue-purple NBT-formazan precipitate at sites of enzyme activity. |
| Detection Method | Colorimetric (chromogenic) detection; dark blue to blue-purple precipitate observed by light microscopy. |
| Sample Type | Cells (e.g., iPSCs, differentiated osteoblasts in 24-well plates), tissue samples, and alkaline phosphatase-conjugated Western blot membranes. |
| Performance Range / Specifications | Staining intensity reflects the level of alkaline phosphatase; minor quantitative differences may not be distinguishable by eye and can be evaluated by grayscale densitometry (with caution regarding result stability, similar to Western blot band quantification). |
| Specificity | Detects alkaline phosphatase activity; suitable for endogenous ALP in cells and tissues and for ALP conjugates on Western blot membranes. |
| Reaction Conditions / Protocol | 1. Prepare the BCIP/NBT staining working solution fresh before use (the working solution must be freshly prepared and cannot be reused). 2. Fix and wash the cell or tissue sample, or use an ALP-conjugated membrane after Western blotting. 3. Add the working solution and incubate until a dark blue to blue-purple color develops at sites of ALP activity. 4. Stop the reaction and observe by light microscopy or image the membrane. |
| Components / Formulation | BCIP staining solution and NBT staining solution (plus associated buffers per kit configuration). |
| Storage Conditions | Store at 4 °C. BCIP staining solution and NBT staining solution must be stored protected from light. |
| Shelf Life | 1 year when stored at 4 °C. |
| Package Specifications | 100 mL |
| Product Form | Liquid kit components. |
| Shipping Conditions | Shipped with ice packs. |
| Expiration Date / Stability | 1 year at 4 °C; the prepared BCIP/NBT working solution must be used fresh and cannot be stored or reused. |
| Compatibility | Suitable for cell and tissue staining (including iPSC identification and osteoblast differentiation assays in multi-well formats such as 24-well plates) and for ALP-based Western blot chromogenic detection. |
| Application Notes / Precautions | The BCIP/NBT staining working solution must be freshly prepared before each use and cannot be reused. Protect BCIP and NBT solutions from light. This kit is not suitable for cell viability assays (use a dedicated viability assay such as a CCK-8 type kit instead). Staining intensity reflects ALP content; subtle differences may require grayscale densitometry. |
For research use only, not for clinical use.
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