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| Product Name | RIPA Lysis Buffer with Protease Inhibitor Cocktail |
| Catalog No. | PTR-HMM-0058 |
| Description | A complete cell and tissue lysis buffer formulated for efficient protein extraction. The buffer contains a balanced mixture of ionic and non-ionic detergents combined with a broad-spectrum protease inhibitor cocktail to maintain protein integrity during extraction. |
| Intended Use | Efficient extraction of total cellular protein from cultured mammalian cells and tissue samples for downstream applications including Western blotting, immunoprecipitation, and protein quantification assays. |
| Principle / Technology | Combined action of sodium deoxycholate (ionic detergent), NP-40 (non-ionic detergent), and SDS (denaturing detergent) to solubilize membrane proteins, cytoplasmic proteins, and nuclear proteins in a Tris-buffered saline system. |
| Detection Method | Mechanical lysis with pipetting or homogenization; protein recovery by centrifugation |
| Sample Type | Cultured adherent and suspension cells, fresh or frozen tissue samples; compatible with most mammalian cell lines |
| Performance Range / Specifications | Typical protein yield: 1–5 mg/mL from 10^7 cultured cells; protein concentration can be determined by BCA or Bradford assay |
| Sensitivity / LOD | Extracts proteins at concentrations as low as 50 μg/mL with proper processing |
| Specificity | Broad protein extraction profile encompassing cytoplasmic, membrane, and nuclear proteins; protease inhibitor cocktail inhibits serine, cysteine, and aspartic acid proteases |
| Reaction Conditions / Protocol | Wash cells with cold PBS, add RIPA buffer (1 mL per 10^7 cells), incubate on ice for 15–30 minutes with intermittent vortexing, centrifuge at 14,000g for 15 minutes at 4°C, collect supernatant |
| Components / Formulation | Ready-to-use RIPA buffer (50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1% NP-40, 0.5% sodium deoxycholate, 0.1% SDS), protease inhibitor cocktail (AEBSF, aprotinin, bestatin, E-64, leupeptin, pepstatin A), EDTA (1 mM) |
| Storage Conditions | 2–8°C; protease inhibitor cocktail is stable for 12 months after addition to buffer; aliquot to avoid repeated freeze-thaw cycles |
| Shelf Life | 18 months for buffer base; 12 months after addition of protease inhibitors |
| Package Specifications | 100 mL (sufficient for approximately 100 extractions from 10^7 cells each) |
| Product Form | Ready-to-use liquid buffer with protease inhibitors pre-added |
| Quality Control | Each lot validated by Western blot with housekeeping protein antibodies (β-actin, GAPDH); tested for protease inhibition efficiency using fluorogenic peptide substrates |
| Key Features | Complete lysis solution eliminates need for separate protease inhibitor addition; optimized detergent ratio for maximum protein solubilization; suitable for direct use in BCA and Bradford protein assays |
For research use only, not for clinical use.
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