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| Product Name | Senescence-Associated β-Galactosidase Staining Kit |
| Catalog No. | CCAT-HMM-0057 |
| Description | A histochemical staining kit for detecting senescence-associated beta-galactosidase activity at pH 6.0 in cultured cells and tissue sections. Senescent cells express a lysosomal β-galactosidase isoform active at pH 6.0 that is not detectable in quiescent or proliferating cells under the same conditions. The chromogenic X-Gal substrate produces an insoluble blue precipitate localized within senescent cells, enabling direct visualization by light microscopy. |
| Intended Use | Identification and visualization of senescent cells in culture and tissue by chromogenic detection of senescence-associated β-galactosidase. |
| Principle / Technology | Chromogenic X-Gal cleavage by senescence-associated β-galactosidase at pH 6.0 producing localized blue precipitate |
| Detection Method | Light microscopy (brightfield) |
| Sample Type | Adherent cultured cells, frozen tissue sections, or fresh tissue biopsies |
| Performance Range / Specifications | Incubation time: 12–16 hr at 37 °C (without CO₂); blue staining visible in senescent cells from 2 hr onward |
| Sensitivity / LOD | Detects senescence in as few as 5% of cells in a mixed population |
| Specificity | β-galactosidase activity at pH 6.0 distinguishes senescent cells from lysosomal β-galactosidase active at pH 4.0 |
| Reaction Conditions / Protocol | Wash cells with PBS; fix with 0.5% glutaraldehyde 10 min; wash 2×; add staining solution (X-Gal in citric acid/sodium phosphate buffer, pH 6.0, with potassium ferrocyanide and potassium ferricyanide) containing X-Gal, incubate 12–16 hr at 37 °C; observe by microscopy |
| Components / Formulation | X-Gal chromogenic substrate (20 mg/mL in DMF), citric acid/sodium phosphate buffer (pH 6.0), potassium ferrocyanide, potassium ferricyanide, glutaraldehyde fixative solution, PBS |
| Storage Conditions | X-Gal: –20 °C protected from light; buffer: room temperature; fixative: 2–8 °C |
| Shelf Life | 12 months for X-Gal; 24 months for buffers |
| Package Specifications | 100 staining reactions (35 mm dishes) per kit |
| Product Form | Liquid solutions; X-Gal as DMF solution |
| Quality Control | Functionally validated with senescent human dermal fibroblasts (passage ≥15 vs passage 5 negative control) |
| Key Features | pH 6.0 specificity provides clean distinction between senescent and non-senescent cells in mixed populations |
| Purity | Reagent grade; fluorophore-labeled reagents ≥90% labeling efficiency |
| Concentration | As specified per kit; sufficient for stated number of tests |
| Activity / Unit Definition | Apoptosis induction confirmed by positive control treatment producing ≥5-fold signal increase |
| Molecular Weight | Varies by dye and enzyme component |
| Source / Origin | Recombinant annexin proteins; synthetic fluorophore conjugates; chromogenic/fluorogenic peptide substrates |
| pH Range / Optimal pH | Binding/washing buffer pH 7.2–7.4 |
| Shipping Conditions | Cold packs; protect fluorophores from light; some components may require –20 °C |
| Expiration Date / Stability | 6–12 months at recommended storage temperature |
| Regulatory / Compliance | Research use; manufactured under ISO 9001 |
| Compatibility | Compatible with flow cytometry and fluorescence microscopy platforms |
| Recommended Buffer System | Annexin binding buffer with calcium; cell lysis buffer with protease inhibitors for caspase assays |
| Application Notes / Precautions | Process samples promptly after staining; include unstained and single-stain controls for flow cytometry compensation; use appropriate apoptosis inducers as positive controls |
| Batch-to-Batch Consistency | Positive control signal within ±20% of reference lot; staining pattern consistent |
For research use only, not for clinical use.
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