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| Product Name | Alizarin Red S Calcium Staining Kit for Mineralized Tissue |
| Catalog No. | ATRSA-0028 |
| Description | Histochemical staining kit using Alizarin Red S for detection and quantification of calcium deposits in tissue sections and cell cultures. Calcium ions form an Alizarin Red S-calcium chelate complex producing bright orange-red staining at sites of mineralization. Essential for bone histomorphometry, osteogenic differentiation assessment, vascular calcification studies, and kidney stone analysis. |
| Intended Use | Detection of calcium phosphate and calcium carbonate deposits in tissue sections; osteogenic differentiation assessment in cell cultures; vascular and valvular calcification studies; kidney stone composition analysis; bone tissue engineering scaffold mineralization evaluation; dental pulp calcification detection. |
| Principle / Technology | Alizarin Red S (1,2-dihydroxyanthraquinone-3-sulfonic acid sodium salt) chelates calcium ions at pH 4.1-4.3 to form an insoluble Alizarin Red S-calcium complex (birefringent orange-red); staining is pH-dependent — acidic pH provides specificity for calcium over magnesium and other cations |
| Detection Method | For tissue: deparaffinize, hydrate; stain in Alizarin Red S (pH 4.2) 5-30 min (monitor); rinse in distilled water; dehydrate quickly through acetone and acetone-xylene; clear and mount. For cell culture: fix with 4% PFA 15 min; stain 30 min; wash; visualize |
| Sample Type | FFPE tissue sections (bone, artery, heart valve, kidney); undecalcified bone sections (plastic-embedded); cell culture monolayers (osteogenic differentiated MSCs, osteoblasts) |
| Sensitivity / LOD | Detects calcium deposits as small as single calcium phosphate nodules; visual detection limit ~0.1 µg calcium per spot |
| Specificity | Specific for calcium under pH-controlled conditions; magnesium and other divalent cations produce minimal staining at pH 4.1-4.3; phosphate alone does not stain; iron deposits may produce false-positive — use Prussian blue to confirm |
| Reaction Conditions / Protocol | Tissue sections: 5-30 min RT; cell cultures: 30 min RT; differentiation important — overstaining causes non-specific background |
| Components / Formulation | Alizarin Red S staining solution (2%, pH 4.1-4.3, filtered), positive control slide (calcified artery or bone), detailed protocol for tissue and cell culture applications |
| Storage Conditions | 15-25 °C; protect from light; tightly sealed to prevent pH drift |
| Shelf Life | 24 months from manufacture date |
| Package Specifications | Kit for 50 slides, 100 slides |
| Product Form | Liquid reagent, ready-to-use |
| Key Features | pH-optimized for calcium specificity (pH 4.2); ready-to-use — no daily preparation; includes positive control; protocol for both tissue and cell culture; quantification by cetylpyridinium chloride extraction method possible; compatible with brightfield microscopy |
| Purity | Alizarin Red S certified by Biological Stain Commission; dye content ≥65%; pH verified per lot at 4.2 ± 0.1 |
| Concentration | As specified; ready-to-use or concentrate format |
| Activity / Unit Definition | Staining intensity and specificity validated on reference tissue/cell preparations |
| Molecular Weight | Varies by dye component as specified in product documentation |
| Source / Origin | Synthetic dyes and biological stains; certified by Biological Stain Commission where applicable |
| pH Range / Optimal pH | pH 6.0–7.0 for Romanowsky-type stains; varies by formulation |
| Shipping Conditions | Ambient temperature |
| Expiration Date / Stability | 24 months at 15-25 °C; discard if solution pH drifts outside 4.1-4.3 range or if heavy precipitate forms; filter before use if slight precipitate observed |
| Regulatory / Compliance | For laboratory and research use only; RUO; Alizarin Red S is a laboratory chemical — avoid skin contact and inhalation; ISO 9001 |
| Compatibility | Compatible with FFPE and frozen sections; undecalcified bone sections give strongest signal; EDTA-decalcified bone may show reduced staining due to calcium removal — use acid-decalcified controls; for cell culture quantification, dissolve bound dye with 10% cetylpyridinium chloride and measure OD at 562 nm |
| Recommended Buffer System | Use with provided buffer or phosphate buffer pH 6.8 as specified |
| Application Notes / Precautions | Avoid contact with skin and clothing; stains are permanent; use appropriate PPE |
| Batch-to-Batch Consistency | Staining intensity and differential contrast validated on reference specimens per lot |
For research use only, not for clinical use.
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