ReadyProbes™ Reagent F-Actin Phalloidin Conjugates

ActinGreen 488 ReadyProbes Reagent is a selective, high-affinity F-actin probe conjugated to our bright, photostable, green-fluorescent Alexa Fluor 488 dye. ActinGreen 488 reagent is a room temperature-stable solution of Alexa Fluor 488 Phalloidin (Cat. No. A12379) packaged in a dropper bottle. ActinRed 555 ReadyProbes Reagent is a comparable F-actin stain conjugated to the red-orange fluorescent dye tetramethylrhodamine (TRITC). ActinRed 555 reagent is a room temperature-stable solution of Rhodamine Phalloidin (Cat. No. R415) packaged in a dropper bottle.

ActinRed 568 ReadyProbes Reagent is our high-affinity phalloidin probe conjugated to our bright, red-fluorescent Alexa Fluor 568 dye. This dye is packaged in an easy to use dropper-optional bottle, allowing end-users to choose either to pipet out of the tube using the classic cap, or utilize the dropper bottle tip for typical ReadyProbes workflows.

ActinRed 647 ReadyProbes Reagent leverages our high-affinity phalloidin conjugated to our bright Far-Red Alexa Fluor 647 Plus dye, offering enhanced brightness vs. classic Alexa Fluor 647 phalloidin conjugates, in an easy to use format. ActinRed 647 ReadyProbes Reagent should be stored from 2-8°C for long-term, and also offers the option to leverage pipetting, if desired, via the dropper-optional cap.

These ready-to-use solutions combine superior brightness with exceptional F-actin staining. Phalloidin is a bi-cyclic peptide, commonly used in imaging applications to selectively label F-actin. Fluorescently labeled phalloidin has several advantages over antibodies for actin labeling, including virtually identical binding properties with actin from different species of plants and animals and low non-specific binding.

Features

• High-affinity staining of F-actin with superior specificity compared to antibody methods
• ActinGreen 488 Excitation/Emission: (495/518 nm), detected through standard GFP and FITC filter sets
• ActinRed 555 Excitation/Emission: (540/565 nm), detected through standard RFP and TRITC filter sets
• ActinRed 568 Excitation/Emission: (568/600 nm), detected through standard Texas Red filter sets
• ActinRed 647 Excitation/Emission: (647/660 nm), detected through standard Cy5 filter sets
• Use counterstain (i.e., DAPI or NucBlue ReadyProbes reagent) as desired

ReadyProbes format advantages

• Ready-to-use formulation—no need to dilute, weigh, or pipette
• Just use two drops per mL of medium to stain, incubate 30 minutes, wash, then image

Applications

• For fixed and permeabilized cells or tissue
• Fluorescence microscopy (IF/IHC/ICC)
• High content screening, high content imaging

Suggestions for use

• Both ActinRed and ActinGreen reagents may be added directly to fixed cells in full medium or buffer solutions
• In most cases 2 drops/mL and an incubation time of 15–30 minutes is sufficient for bright actin staining; however, optimization may be needed for some cell types, conditions, and applications. In such cases, simply add more or fewer drops until the optimal staining intensity is obtained.