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| Product Name | NADP/NADPH Quantification Fluorometric Assay Kit |
| Catalog No. | CMTR-HMM-0073 |
| Description | Fluorometric assay for separate quantification of NADP⁺ and NADPH in cell and tissue lysates using specific enzymatic cycling. Selective acid/base extraction protocols differentially preserve oxidized and reduced forms, enabling NADP⁺/NADPH ratio calculation — a critical indicator of cellular redox state, pentose phosphate pathway activity, and biosynthetic capacity. |
| Intended Use | NADP⁺/NADPH quantification and ratio determination; cellular redox state assessment; oxidative stress studies; pentose phosphate pathway activity analysis; NADPH-dependent antioxidant system evaluation. |
| Principle / Technology | NADP⁺-specific cycling: NADP⁺ reduced to NADPH, which reduces fluorogenic probe; cycling amplifies 100-1,000×; selective extraction separates NADP⁺ from NADPH |
| Detection Method | Fluorometric; Ex/Em 530/590 nm; colorimetric backup at 570 nm |
| Sample Type | Cell lysates (10⁵-10⁶ cells), tissue homogenates (10-50 mg); separate extraction for each form |
| Sensitivity / LOD | LOD 1 nM NADP⁺/NADPH in assay; linear 1-200 nM |
| Specificity | NADP⁺/NADPH specific; NAD⁺/NADH cross-reactivity <0.1% |
| Reaction Conditions / Protocol | Acid extract for total NADP, alkaline extract for NADPH (60 °C, 5 min); neutralize; add cycling mix; read fluorescence after 30 min |
| Components / Formulation | NADP cycling enzyme mix, NADPH standard (10 nmol), fluorogenic probe, acid extraction buffer, alkaline extraction buffer, neutralization buffer |
| Storage Conditions | Store at -20 °C; protect probe and standards from light |
| Shelf Life | 12 months at -20 °C |
| Package Specifications | 100 assays, 200 assays for full NADP⁺/NADPH determination |
| Product Form | Lyophilized enzyme and standard; liquid buffers |
| Key Features | Separate NADP⁺/NADPH quantification; enzymatic cycling amplification; selective extraction protocols; NADPH standard included; 96-well format |
| Purity | NADPH standard >98% HPLC; cycling enzyme >200 U/mg; no NAD⁺/NADH contamination |
| Concentration | As specified per kit format; probe stock solutions in DMSO as specified |
| Activity / Unit Definition | Standard curve linearity R² ≥0.99 per lot; dynamic range as specified |
| Molecular Weight | Varies by probe and buffer component |
| Source / Origin | Synthetic fluorescent probes; recombinant enzymes; purified metabolite standards |
| pH Range / Optimal pH | pH 7.2–7.4 for cell-based assays; pH varies for biochemical metabolite detection |
| Shipping Conditions | Cold pack -20 °C; standards and probe light-sensitive |
| Expiration Date / Stability | 12 months at -20 °C; reconstituted standard 2 weeks at 4 °C |
| Regulatory / Compliance | For laboratory and research use only; RUO; ISO 9001 |
| Compatibility | Compatible with fluorescence microplate readers, luminometers, and spectrophotometers |
| Recommended Buffer System | Assay-specific buffer optimized per product protocol |
| Application Notes / Precautions | Normalize signal to cell number or protein content; avoid probe photobleaching; include appropriate antioxidant controls for ROS assays |
| Batch-to-Batch Consistency | Standard curve parameters within ±15% of reference lot |
For research use only, not for clinical use.
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