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| Product Name | Cell Lysis Diluent for Chemiluminescent Immunoassay, Non-Denaturing |
| Catalog No. | ATRD-0028 |
| Description | Non-denaturing diluent specifically designed for chemiluminescent immunoassay (CLIA) sample preparation involving cell lysates, tissue homogenates, and complex biological matrices. The gentle nonionic detergent and protein stabilizer system extracts and dilutes intracellular antigens while preserving native protein conformation and antibody epitope accessibility. Formulated without denaturing agents (SDS, urea, guanidine) to maintain compatibility with sandwich and competitive CLIA formats using acridinium ester or alkaline phosphatase-AMPPD detection systems. |
| Intended Use | Lysis of cultured cells and dilution of tissue homogenates for quantitative chemiluminescent immunoassay detection of intracellular proteins, phosphorylated signaling molecules, and cytoplasmic biomarkers on automated CLIA platforms. |
| Principle / Technology | Nonionic detergent (NP-40 alternative) gently permeabilizes cell membranes without denaturing protein structure; protein stabilizers (trehalose, BSA) maintain antigen conformation during lysis; protease/phosphatase inhibitor cocktail preserves post-translational modifications. |
| Detection Method | Add 100-200 µL diluent per 1×10^6 pelleted cells; pipette to resuspend; incubate 15 minutes on ice with periodic vortexing; centrifuge at 14,000 × g for 10 minutes at 4 °C; collect supernatant for CLIA analysis. |
| Sample Type | Cultured adherent and suspension cells, fresh and frozen tissue homogenates, immunoprecipitated protein complexes. |
| Performance Range / Specifications | Protein extraction efficiency >90% compared to RIPA buffer as measured by total protein assay; preserves >85% of native protein structure as assessed by circular dichroism. |
| Sensitivity / LOD | Detectable CLIA signal for low-abundance antigens down to 0.5 pg/mL with appropriate antibody pair; linear dynamic range 0.5-10,000 pg/mL. |
| Specificity | Non-denaturing extraction preserves native epitope structure; does not interfere with acridinium ester chemiluminescence or alkaline phosphatase enzyme activity. |
| Reaction Conditions / Protocol | Incubate cells with diluent for 15 minutes on ice; vortex every 5 minutes; centrifuge at 14,000 × g, 4 °C, 10 minutes; use supernatant directly or dilute further in CLIA assay buffer. |
| Components / Formulation | Nonionic detergent blend (0.5% v/v), trehalose 5% w/v, BSA 0.1% w/v, HEPES 50 mM pH 7.4, NaCl 150 mM, EDTA 1 mM, protease inhibitor cocktail (AEBSF, aprotinin, bestatin, E-64, leupeptin, pepstatin A), phosphatase inhibitor cocktail (sodium orthovanadate, sodium pyrophosphate, β-glycerophosphate). |
| Storage Conditions | Store at -20 °C; avoid repeated freeze-thaw cycles; thaw on ice before use. |
| Shelf Life | 12 months from date of manufacture at -20 °C. |
| Package Specifications | 10 mL, 25 mL, 50 mL bottles; ready-to-use with inhibitors pre-added. |
| Product Form | Clear, colorless to pale yellow liquid; sterile-filtered 0.22 µm. |
| Quality Control | Each lot tested for protein extraction efficiency using standardized HeLa cell pellet, CLIA interference testing, protease and phosphatase inhibitor activity verification, and sterility. |
| Key Features | Non-denaturing detergent system; pre-formulated with protease and phosphatase inhibitors; protein stabilizers included (trehalose, BSA); compatible with acridinium ester and ALP-AMPPD CLIA detection; preserves phospho-epitope immunoreactivity. |
| Purity | Detergent blend H2O2-free certified; BSA endotoxin <0.1 EU/mg; trehalose ≥99% purity. |
| Concentration | Ready-to-use 1× formulation; inhibitor concentrations optimized for mammalian cell extracts. |
| Activity / Unit Definition | Protease inhibitor activity validated by complete inhibition of trypsin (100 µg/mL) for >2 hours at 4 °C. |
| Molecular Weight | Not applicable — complex multi-component formulation. |
| Source / Origin | Synthetic detergents, recombinant protein stabilizers, analytical grade buffer salts and inhibitors. |
| pH Range / Optimal pH | pH 7.4 ± 0.1 at 25 °C. |
| Shipping Conditions | Dry ice (-80 °C) recommended for long-distance transit; cold pack acceptable for overnight delivery. |
| Expiration Date / Stability | 12 months at -20 °C; after thawing, store at 2-8 °C and use within 1 week; do not refreeze. |
| Regulatory / Compliance | For research use only; not for diagnostic procedures. Manufactured in ISO 9001 certified facility. |
| Compatibility | Compatible with acridinium ester, isoluminol, and alkaline phosphatase-AMPPD (adamantyl 1,2-dioxetane aryl phosphate) chemiluminescent detection systems. Compatible with magnetic bead-based and microplate CLIA formats. Nonionic detergent does not interfere with antibody-antigen binding kinetics. |
| Recommended Buffer System | 50 mM HEPES, 150 mM NaCl, 1 mM EDTA, pH 7.4. |
| Application Notes / Precautions | Pre-chill diluent and all tubes on ice before use. Add fresh PMSF (1 mM final) immediately before use if serine protease activity is a concern (PMSF has short half-life in aqueous solution). For phospho-protein analysis, verify phosphatase inhibitor cocktail activity with a phospho-standard before critical experiments. Avoid over-vortexing as foam formation may denature sensitive proteins. |
| Batch-to-Batch Consistency | Protein extraction yield within ±10% of reference lot using standardized HeLa cell input; inhibitor activity verified per lot. |
For research use only, not for clinical use.
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