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| Product Name | Fluor 750 Phalloidin |
| Catalog No. | RESK-0580 |
| Description | Fluor 750 Phalloidin is a near-infrared fluorescent conjugate of phalloidin, a cyclic heptapeptide toxin derived from the mushroom Amanita phalloides. Phalloidin selectively binds filamentous actin (F-actin) with high affinity (Kd = 20 nM) and does not bind monomeric actin (G-actin), making it a standard tool for labeling F-actin in tissue sections, cell cultures, and cell-free systems for qualitative and quantitative analysis. Phalloidin conjugates bind large and small actin filaments in muscle and non-muscle cells of both animal and plant origin with approximately one phalloidin molecule per actin subunit, and nonspecific binding is negligible, providing exceptionally sharp contrast between stained and unstained regions. Phalloidin derivatives are very small (about 12-15 Å in diameter, molecular weight < 2000 Da), so many physiological properties of the labeled actin are preserved: actin-binding proteins such as myosin, tropomyosin, and DNase I can still interact with labeled filaments, phalloidin-labeled filaments can still penetrate solid-phase myosin matrices, and glycerol-extracted muscle fibers remain contractile after labeling. Phalloidin binding also prevents the dissociation of F-actin (microfilaments), stabilizes microfilament structure, and disrupts the polymerization-depolymerization equilibrium, lowering the critical concentration (CC) of actin polymerization to < 1 µg/mL; phalloidin therefore also acts as a polymerization promoter and inhibits the ATP-hydrolyzing activity of F-actin. Fluor 750 Phalloidin delivers highly specific staining with strong contrast, outperforming actin antibodies for qualitative and quantitative F-actin detection. Fluor 750 Phalloidin staining is fully compatible with other fluorescent labels used in cell analysis, including fluorescent proteins, semiconductor nanocrystals, and other Fluor dye conjugates (including Fluor-conjugated secondary antibodies). F-actin bound by this product retains many of the native biological properties of actin, and binding shows no species specificity, ensuring broad applicability. The product is supplied as powder pellets; reconstitute with 30 µL DMSO and mix thoroughly before use. |
| Intended Use | For fluorescent labeling of filamentous actin (F-actin) in tissue sections, cultured cells, and cell-free systems, enabling qualitative and quantitative analysis of the actin cytoskeleton. |
| Principle / Technology | Phalloidin is a cyclic heptapeptide toxin from Amanita phalloides that binds F-actin selectively and with high affinity (Kd = 20 nM) without binding G-actin. Conjugation to the near-infrared fluorescent dye Fluor 750 allows direct visualization of F-actin. Binding stabilizes microfilaments by blocking their dissociation, lowers the critical concentration of actin polymerization to < 1 µg/mL, and inhibits F-actin ATP hydrolysis. |
| Detection Method | Fluorescence microscopy / fluorescence imaging |
| Sample Type | Tissue sections, cultured cells, and cell-free systems; muscle and non-muscle cells of animal or plant origin |
| Performance Range / Specifications | Size: 300 T Maximum excitation/emission (Ex/Em): 757/779 nm Molecular weight: ~3300 Solubility: soluble in DMSO |
| Sensitivity / LOD | High-affinity binding to F-actin with Kd = 20 nM; nonspecific binding is negligible, giving clearly distinguishable stained and unstained regions. |
| Specificity | Selectively binds filamentous actin (F-actin); does not bind monomeric actin (G-actin). No species-dependent differences in binding. |
| Reaction Conditions / Protocol | 1. Supplied as powder pellets; add 30 µL DMSO and mix thoroughly to prepare the stock solution. 2. Fix cells with freshly prepared 4% paraformaldehyde in PBS; avoid methanol-containing fixatives, as methanol can disrupt F-actin during fixation. 3. Permeabilize cells adequately; extend permeabilization time if staining is weak or incomplete. 4. Apply enough working solution to completely cover the cells; optimize the working concentration and staining time for each cell type. 5. For imaging, locate fields with low-energy laser excitation and switch to higher energy only for observation and acquisition to minimize photobleaching. |
| Components / Formulation | Fluor 750 Phalloidin (Fluor 750-conjugated phalloidin), supplied as powder pellets; soluble in DMSO. |
| Storage Conditions | Store at -25 to -15 °C. |
| Shelf Life | 1 year |
| Package Specifications | 300 T |
| Product Form | Powder pellets |
| Molecular Weight | ~3300 |
| Source / Origin | Phalloidin derived from the mushroom Amanita phalloides |
| Shipping Conditions | Shipped on ice packs. |
| Expiration Date / Stability | Valid for 1 year when stored at -25 to -15 °C. |
| Regulatory / Compliance | For research use only. |
| Compatibility | Fully compatible with other fluorescent stains used in cell analysis, including fluorescent proteins, semiconductor nanocrystals, and other Fluor dye conjugates (including Fluor-conjugated secondary antibodies). F-actin bound by this product retains many native biological properties of actin. Binding is species-independent. |
| Recommended Buffer System | PBS (e.g., 4% paraformaldehyde in PBS for fixation) |
| Application Notes / Precautions | 1. Rapid fluorescence fading after excitation may result from excessive reagent concentration; reduce the working concentration and use low-energy laser excitation when searching for fields. 2. Do not co-stain with click chemistry-based labeling reactions (e.g., EdU), which can abolish the phalloidin fluorescence signal. 3. Avoid methanol-containing fixatives, as methanol may disrupt F-actin during fixation. 4. Solvents such as xylene or acetone (e.g., during paraffin section deparaffinization) prevent phalloidin binding to F-actin; use cryosections (which generally do not require organic solvent washing) instead of paraffin sections, or switch to an anti-actin antibody. 5. If staining is uneven or weak, optimize the working concentration and staining time, fix with freshly prepared 4% paraformaldehyde, and extend permeabilization time if needed. |
| Recognition Site | Filamentous actin (F-actin); binds approximately one phalloidin molecule per actin subunit |
For research use only, not for clinical use.
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