- Home
- IVD
- By Technology Types
- By Diseases Types
- By Product Types
- Research
- Resource
- Distributors
- Company
| Product Name | Intracellular Calcium Flux Detection Kit (Fluo-4 AM) |
| Catalog No. | CMTR-HMM-0061 |
| Description | A fluorescent intracellular calcium indicator kit using Fluo-4 AM, a cell-permeable acetoxymethyl ester derivative of the calcium-sensitive dye Fluo-4. Fluo-4 AM passively enters cells, is cleaved by intracellular esterases to release the active Fluo-4 indicator, which undergoes a dramatic fluorescence enhancement (>100-fold) upon calcium binding. |
| Intended Use | Intracellular calcium measurement in live cells for studies of G protein-coupled receptor signaling, ion channel function, neuronal activity, muscle contraction, immune cell activation, and calcium-mediated signal transduction pathways. |
| Principle / Technology | Fluo-4 AM contains acetoxymethyl ester groups that mask the calcium-binding tetracarboxylate moiety, enabling passive diffusion across the plasma membrane; intracellular esterases hydrolyze the AM esters to release the active Fluo-4 indicator, which is trapped inside the cell as a polyanionic species; upon binding free cytosolic Ca2+ (Kd approximately 335 nM), Fluo-4 undergoes a conformational change resulting in >100-fold fluorescence enhancement at Ex/Em 494/516 nm. |
| Detection Method | Fluorescence microscopy with FITC/GFP filter set (Ex/Em 494/516 nm), flow cytometry (488 nm laser, FL1 channel), or fluorescence plate reader; confocal microscopy for calcium localization |
| Sample Type | Live adherent and suspension mammalian cells, primary neurons and astrocytes, cardiomyocytes, immune cells (T cells, B cells, macrophages), cells expressing recombinant GPCRs or calcium channels |
| Performance Range / Specifications | Fluo-4 AM loading concentration: 1–5 μM; loading time: 15–60 minutes at 37°C; dynamic range typically 5–20× fluorescence increase from resting (approximately 100 nM Ca2+) to stimulated (1–2 μM Ca2+) conditions; Kd for calcium approximately 335 nM |
| Sensitivity / LOD | Detects intracellular calcium transients with amplitudes as low as 50 nM above resting levels; single-cell resolution by fluorescence microscopy and flow cytometry |
| Specificity | Fluo-4 specifically binds Ca2+ with 5–10× greater brightness than Fluo-3 in response to equivalent calcium changes; affinity for Mg2+ and other divalent cations is negligible under physiological conditions; intracellular retention depends on active esterase cleavage and is reversible by probenecid transporter inhibition |
| Reaction Conditions / Protocol | Prepare 2–5 mM Fluo-4 AM stock in anhydrous DMSO; dilute to 1–5 μM in serum-free medium or HBSS containing 0.02% Pluronic F-127 (dispersion aid); add loading solution to cells; incubate 30 minutes at 37°C (or 45–60 minutes at room temperature); wash cells with indicator-free buffer; optionally add probenecid (2.5 mM) to inhibit dye extrusion by organic anion transporters; incubate additional 30 minutes to allow intracellular esterase hydrolysis; stimulate cells with agonist or depolarizing stimulus; acquire fluorescence time-course data |
| Components / Formulation | Fluo-4 AM (lyophilized, 50 μg or 1 mg vials, or 1 mM DMSO stock), Pluronic F-127 (20% solution in DMSO), probenecid (water-soluble sodium salt), detailed loading and calibration protocol including instructions for Ca2+ calibration using ionomycin and EGTA |
| Storage Conditions | Fluo-4 AM lyophilized: -20°C protected from light and moisture, stable 24 months; DMSO stock (1–5 mM): -20°C protected from light, desiccated, stable 3 months; working loading solution: prepare fresh, use within 2 hours |
| Shelf Life | 24 months from date of manufacture for lyophilized dye |
| Package Specifications | 20 × 50 μg vials, 1 mg vial, or 10 × 50 μg special packaging for plate reader format |
| Product Form | Lyophilized dye in amber glass vials or DMSO stock solution |
| Quality Control | Each lot tested for fluorescence enhancement upon calcium binding using Ca2+ calibration buffers (0–39 μM free Ca2+); purity ≥95% by HPLC; residual AM ester hydrolysis confirmed complete within 30 minutes in HeLa cell loading assay |
| Key Features | Enhanced brightness compared to Fluo-3 and Calcium Green-1; excitation optimum at 488 nm compatible with argon-ion lasers; single-wavelength excitation ratio provides simplified optical setup; Pluronic F-127 inclusion improves dye solubility and cellular loading efficiency |
For research use only, not for clinical use.
|
There is no product in your cart. |
Copyright © 2026 Alta DiagnoTech. All rights reserved.
