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| Product Name | Fluor 647 Phalloidin |
| Catalog No. | RESK-0557 |
| Description | Phalloidin is a cyclic heptapeptide toxin derived from the death cap mushroom (Amanita phalloides). It selectively binds filamentous actin (F-actin) with high affinity (Kd = 20 nM) and does not bind monomeric actin (G-actin). It is commonly used to label F-actin in tissue sections, cell cultures, or cell-free systems for qualitative and quantitative analysis of F-actin. Phalloidin derivatives bind large and small filaments with similar affinity in both muscle and non-muscle cells of animal and plant origin, at a stoichiometry of approximately one phalloidin molecule per actin subunit. Nonspecific binding is virtually negligible, giving a very clear distinction between stained and unstained regions, so phalloidin derivatives are particularly suitable as a replacement for actin antibodies in related research. Phalloidin derivatives are very small (about 12-15 Angstrom in diameter, molecular weight below 2000 Daltons), so many physiological properties of the labeled actin are preserved: actin-binding proteins such as myosin, tropomyosin, and DNase I can still react; phalloidin-labeled filaments can still penetrate solid-phase myosin matrices; and labeled glycerol-extracted myofibrils can still contract. Binding of phalloidin prevents depolymerization of filamentous actin (microfilaments) and stabilizes the microfilament structure, disrupting the polymerization-depolymerization equilibrium. This lowers the critical concentration (CC) for actin polymerization to below 1 μg/mL, so phalloidin can also serve as a polymerization promoter. In addition, phalloidin inhibits the ATP hydrolysis activity of F-actin. This product is Fluor 647-labeled phalloidin, offering high brightness and photostability with strong staining specificity and high contrast, providing better staining performance than actin antibodies for qualitative and quantitative F-actin detection. Fluor 647 Phalloidin staining is fully compatible with other fluorescent labels used in cell analysis, including fluorescent proteins, semiconductor nanocrystals, and other fluorescent dye conjugates (including dye-conjugated secondary antibodies). F-actin bound by this product retains many of the biological properties of actin itself, and the binding shows no species difference, giving broad applicability. The product is supplied as a powder; dissolve in 30 uL of DMSO to prepare a 1 mg/mL solution. |
| Intended Use | Qualitative and quantitative fluorescent labeling of filamentous actin (F-actin) in tissue sections, cell cultures, or cell-free systems; an alternative to actin antibodies for cytoskeleton research. |
| Principle / Technology | Phalloidin selectively binds F-actin with high affinity (Kd = 20 nM) at a stoichiometry of approximately one phalloidin molecule per actin subunit, without binding G-actin. Binding stabilizes microfilaments, blocks depolymerization, lowers the critical concentration for actin polymerization to below 1 μg/mL, and inhibits F-actin ATP hydrolysis. Conjugation to the bright, photostable far-red Fluor 647 fluorophore (Ex/Em 650/665 nm) enables high-contrast fluorescence detection. |
| Detection Method | Far-red fluorescence detection, Ex/Em 650/665 nm. |
| Sample Type | Tissue sections, cultured cells, or cell-free systems; suitable for muscle and non-muscle cells of animal and plant origin; compatible with fixed samples. |
| Performance Range / Specifications | Molecular weight: 1408.65 Maximum excitation/emission (Ex/Em): 650/665 nm Solubility: soluble in DMSO |
| Specificity | Selectively binds filamentous actin (F-actin) with Kd = 20 nM and does not bind monomeric G-actin; nonspecific binding is virtually negligible, giving clear contrast between stained and unstained regions. Binding shows no species difference and is broadly applicable. |
| Reaction Conditions / Protocol | Reconstitute the powder with 30 uL of DMSO to obtain a 1 mg/mL stock solution. For staining of the cytoskeleton in fixed cell samples, both fixation (e.g., 4% paraformaldehyde) and permeabilization (e.g., Triton X-100) should be performed; fixed but unpermeabilized cells may be stained only partially because altered membrane permeability allows incomplete probe entry. |
| Components / Formulation | Fluor 647-labeled phalloidin, supplied as a powder; reconstitute with 30 uL of DMSO to prepare a 1 mg/mL solution. |
| Storage Conditions | Store at -20 °C, dry and protected from light, after receipt. |
| Shelf Life | One year when stored at -20 °C, dry and protected from light. |
| Package Specifications | 300 tests |
| Product Form | Powder |
| Concentration | 1 mg/mL after reconstitution with 30 uL of DMSO |
| Molecular Weight | 1408.65 |
| Shipping Conditions | Shipped with ice packs. |
| Expiration Date / Stability | Valid for one year when stored at -20 °C, dry and protected from light. |
| Regulatory / Compliance | For research use only. |
| Compatibility | Fully compatible with other fluorescent labels used in cell analysis, including fluorescent proteins, semiconductor nanocrystals, and other fluorescent dye conjugates (including dye-conjugated secondary antibodies). |
| Recommended Buffer System | Soluble in DMSO; prepare stock solutions in DMSO. |
| Application Notes / Precautions | Phalloidin derivatives are very small (12-15 Angstrom in diameter, molecular weight below 2000 Daltons), so many physiological properties of labeled actin are retained, including interactions with myosin, tropomyosin, and DNase I, penetration of solid-phase myosin matrices, and contraction of labeled glycerol-extracted myofibrils. Binding stabilizes microfilaments and promotes actin polymerization (critical concentration lowered to below 1 μg/mL); it also inhibits F-actin ATPase activity. For fixed live-cell samples, perform both fixation and permeabilization to achieve complete cytoskeleton staining. |
For research use only, not for clinical use.
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