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iFluor™ Phalloidin Conjugates for Labeling F-actins


Image of HeLa cells. Actin filaments were stained with Phalloidin-iFluor® 680 conjugate, and nuclei were stained with Nuclear Green™ DCS1.
Actin is a globular, roughly 42-kDa protein found in almost all eukaryotic cells. It is also one of the most highly-conserved proteins, differing by no more than 20% in species as diverse as algae and humans. Actin is the monomeric subunit of two types of filaments in cells: microfilaments, one of the three major components of the cytoskeleton, and thin filaments, part of the contractile apparatus in muscle cells. Thus, actin participates in many important cellular processes including muscle contraction, cell motility, cell division and cytokinesis, vesicle and organelle movement, cell signaling, as well as the establishment and maintenance of cell junctions and cell shape. Phalloidin, a bicyclic heptapeptide toxin, binds specifically at the interface between F-actin subunits, locking adjacent subunits together. Phalloidin binds to actin filaments much more tightly than to actin monomers, leading to a decrease in the rate constant for the dissociation of actin subunits from filament ends, essentially stabilizing actin filaments through the prevention of filament depolymerization. Moreover, phalloidin is found to inhibit the ATP hydrolysis activity of F-actin. Thus, phalloidin traps actin monomers in a conformation distinct from G-actin and it stabilizes the structure of F-actin by greatly reducing the rate constant for monomer dissociation.

AAT Bioquest offers a variety of fluorescent phalloidin derivatives with different colors for multicolor imaging of F-actin. Fluorescent derivatives of phalloidin have turned out to be enormously useful in localizing actin filaments in living or fixed cells as well as for visualizing individual actin filaments in vitro. Fluorescent phalloidin derivatives have been used as an important tool in the study of actin networks at high resolution. Used at nanomolar concentrations, phalloidin derivatives are convenient probes for labeling, identifying and quantitating F-actins in formaldehyde-fixed and permeabilized tissue sections, cell cultures or cell-free experiments. Phalloidin binds to actin filaments much more tightly than to actin monomers, leading to a decrease in the rate constant for the dissociation of actin subunits from filament ends, essentially stabilizing actin filaments through the prevention of filament depolymerization. Moreover, phalloidin is found to inhibit the ATP hydrolysis activity of F-actin. Phalloidin functions differently at various concentrations in cells. When introduced into the cytoplasm at low concentrations, phalloidin recruits the less polymerized forms of cytoplasmic actin as well as filamin into stable "islands" of aggregated actin polymers, yet it does not interfere with stress fibers, i.e. thick bundles of microfilaments.
 

Table 1. Ordering Information for Phalloidin-iFluor® Conjugates

Cat No.
Conjugate
Ex/ Em (nm)
Unit Size
Price
23100Phalloidin-AMCA Conjugate 353/442300 Tests$162
23103Phalloidin-California Red Conjugate 583/605300 Tests$162
23101Phalloidin-Fluorescein Conjugate 492/518300 Tests$162
23110Phalloidin-iFluor® 350344/448300 Tests$162
23111 Phalloidin-iFluor® 405402/425300 Tests$162
23115Phalloidin-iFluor® 488491/516300 Tests$162
23116Phalloidin-iFluor® 514527/554300 Tests$162
23117Phalloidin-iFluor® 532543/563300 Tests$162
23119Phalloidin-iFluor® 555556/569300 Tests$162
23122Phalloidin-iFluor® 594587/603300 Tests$162

 

Cell Navigator® F-Actin Labeling Kits


Cell Navigator® F-Actin Labeling Kits are designed to label F-actins in fixed cells. The kits use fluorescent phalloidin conjugates that are selectively bound to F-actins. The fluorescent phalloidin conjugates are high-affinity probes for F-actins. When used at nanomolar concentrations, phallotoxins are convenient probes for labeling, identifying and quantitating F-actins in formaldehyde-fixed and permeabilized tissue sections, cell cultures or cell-free experiments. The kits provide all the essential components with an optimized staining protocol, which is robust requiring minimal hands-on time.


Images of fixed CPA cells stained with Cell Navigator® F-actin Labeling Kit in a 96-well Costar black wall/clear bottom plate A: Label the cells with 1X Phalloidin-iFluor® 350 for 30 minutes only. B: Treat the cells with phalloidin for 10 minutes, then stain them with 1X Phalloidin-iFluor® 350 for 30 minutes.

 

Table 2. F-actin Labeling Kits

Cat No.
Product Name
Ex (nm)
Em (nm)
Unit Size
Price
22660Cell Navigator® F-Actin Labeling Kit *Blue Fluorescence* 353442500 Tests$217
22661Cell Navigator® F-Actin Labeling Kit *Green Fluorescence* 498520500 Tests$217
22663Cell Navigator® F-Actin Labeling Kit *Orange Fluorescence* 546575500 Tests$217
22664Cell Navigator® F-Actin Labeling Kit *Red Fluorescence* 583603500 Tests$217