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Phalloidin Conjugates
Product key features
- Specific F-Actin Binding: Retains high affinity and specificity for filamentous actin in diverse biological systems.
- Bright and Stable Fluorescence: XFD647 offers exceptional signal intensity and photostability for extended imaging.
- Low Background Signal: Minimal nonspecific binding ensures high-contrast actin visualization.
- Versatile Applications: Reliable in fixed tissues, cultured cells, and actin polymerization assays.
- Multiplex Imaging Ready: Compatible with various fluorescent tags and detection systems for co-labeling experiments.
Product description
XFD647 phalloidin is a high-affinity probe for filamentous actin (F-actin), conjugated to XFD647, a bright, photostable, far-red fluorescent dye. XFD647, manufactured by AAT Bioquest, is structurally equivalent to Alexa Fluor™ 647 (ThermoFisher), ensuring comparable performance in fluorescence applications.
Phalloidin, a bicyclic peptide toxin derived from Amanita phalloides (commonly known as the death cap mushroom), is well known for its high specificity and strong binding affinity to F-actin. When conjugated to XFD647, phalloidin enables precise, high-contrast visualization of F-actin structures with minimal background interference. This fluorescent conjugate is highly effective for imaging F-actin in a wide range of biological specimens, including plant and animal tissues, fixed and permeabilized cells, and cell-free systems.
XFD647 phalloidin binds to F-actin with nanomolar affinity, making it an excellent tool for labeling, identifying, and quantifying actin filaments. Its superior photostability and blinking properties make it particularly well suited for super-resolution microscopy techniques such as SIM and STORM. Additionally, XFD647 phalloidin staining is fully compatible with other fluorescent labels, including fluorescent proteins, Qdot nanocrystals, and iFluor®-conjugated secondary antibodies, allowing seamless integration into multiplex fluorescence imaging workflows.
Example protocol
AT A GLANCE
Prepare samples in microplate wells
Remove liquid from samples in the plate
Add XFD647 Phalloidin Conjugate solution (100 μL/well)
Stain the cells at room temperature for 20 to 90 minutes
Wash the cells
Examine the specimen under microscope with Cy3 filter
Warm the vial to room temperature and centrifuge briefly before opening.
The solution should be stable for at least 6 months if store at -20 °C. Protect the fluorescent conjugates from light, and avoid freeze/thaw cycles.
Note: Phalloidin is toxic, although the amount of toxin present in a vial could be lethal only to a mosquito (LD50 of phalloidin = 2 mg/kg), it should be handled with care.
PREPARATION OF WORKING SOLUTION
Add 1 µL of XFD647 Phalloidin Conjugate solution to 1 mL of PBS with 1% BSA.
Note: The stock solution of phalloidin conjugate should be aliquoted and stored at -20 °C. protected from light.
Note: Different cell types might be stained differently. The concentration of phalloidin conjugate working solution should be prepared accordingly.
SAMPLE EXPERIMENTAL PROTOCOL
Perform formaldehyde fixation. Incubate cells with 3.0–4.0 % formaldehyde in PBS at room temperature for 10–30 minutes.
Note: Avoid any methanol containing fixatives since methanol can disrupt actin during the fixation process. The preferred fixative is methanol-free formaldehyde.
Rinse the fixed cells 2–3 times in PBS.
Optional: Add 0.1% Triton X-100 in PBS into fixed cells for 3 to 5 minutes to increase permeability. Rinse the cells 2–3 times in PBS.
Add 100 μL/well (96-well plate) of XFD647 Phalloidin Conjugate working solution into the fixed cells, and stain the cells at room temperature for 20 to 90 minutes.
Rinse cells gently with PBS 2 to 3 times to remove excess phalloidin conjugate before plating, sealing and imaging under microscope with Cy5 filter set.
Spectrum
Alternative formats
| Name | Conjugate |
| Phalloidin-iFluor® 488 Conjugate | iFluor 488 |
| Phalloidin-iFluor® 647 Conjugate | iFluor 647 |
| Phalloidin-iFluor® 594 Conjugate | iFluor 594 |
| Phalloidin-iFluor® 555 Conjugate | iFluor 555 |
| Phalloidin-iFluor® 633 Conjugate | iFluor 633 |
| Phalloidin-iFluor® 532 Conjugate | iFluor 532 |
| Phalloidin-iFluor® 405 Conjugate | iFluor 405 |
| Phalloidin-iFluor® 514 Conjugate | iFluor 514 |
| Phalloidin-iFluor® 350 Conjugate | iFluor 350 |
Show More (19) | |
Product family
| Name | Excitation (nm) | Emission (nm) | Extinction coefficient (cm -1 M -1) | Quantum yield | Correction Factor (260 nm) | Correction Factor (280 nm) |
| XFD350 Phalloidin *XFD350 Same Structure to Alexa Fluor™ 350* | 343 | 441 | 19000 | - | 0.25 | 0.19 |
| XFD488 Phalloidin *XFD488 Same Structure to Alexa Fluor™ 488* | 499 | 520 | 73000 | 0.921 | 0.3 | 0.11 |
| XFD594 Phalloidin *XFD594 Same Structure to Alexa Fluor™ 594* | 590 | 618 | 92000 | 0.661 | 0.43 | 0.56 |
| XFD647 Azide | 650 | 671 | 270000 | 0.331 | - | 0.03 |
| XFD647 Alkyne | 650 | 671 | 270000 | 0.331 | - | 0.03 |
| XFD647 amine | 650 | 671 | 270000 | 0.331 | - | 0.03 |
| XFD647 acid | 650 | 671 | 270000 | 0.331 | - | 0.03 |
| XFD555 Phalloidin *equivalent to Alexa Fluor® 555 phalloidin* | 553 | 568 | 150000 | 0.11 | 0.08 | 0.08 |
| XFD750 Phalloidin *equivalent to Alexa Fluor® 750 phalloidin* | 752 | 776 | 240000 | 0.121 | 0.00 | 0.04 |
Show More (3) | ||||||
References
Authors: Montecchi-Palmer, Michela and Bermudez, Jaclyn Y and Webber, Hannah C and Patel, Gaurang C and Clark, Abbot F and Mao, Weiming
Journal: Investigative ophthalmology & visual science (2017): 1288-1295
Authors: Muñiz, Javier and Romero, Juan and Holubiec, Mariana and Barreto, George and González, Janneth and Saint-Martin, Madeleine and Blanco, Eduardo and Carlos Cavicchia, Juan and Castilla, Rocío and Capani, Francisco
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Journal: Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society (2014): 239-45
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Journal: Synapse (New York, N.Y.) (2012): 9-19
Authors: Job, Raly and Raja, Vignesh and Grierson, Ian and Currie, Laura and O'Reilly, Steven and Pollock, Natalie and Knight, Emma and Clark, Abbot F
Journal: The British journal of ophthalmology (2010): 1388-92
Safety data sheet