Calculator
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: iFluor® 660 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
Phalloidin iFluor® 660 is an F-actin-specific probe conjugated to the far red-fluorescent dye iFluor® 660. Phalloidin, a bicyclic peptide derived from Amanita phalloides ("death cap" mushroom), is widely used for the selective labeling of filamentous actin (F-actin) in fluorescence microscopy.
The conjugation of phalloidin to iFluor® 660 retains its high binding affinity and specificity for F-actin, providing reliable and reproducible labeling across various biological systems. The iFluor® 660 fluorophore offers exceptional brightness and photostability, enabling prolonged imaging sessions with minimal photobleaching. Additionally, the probe exhibits negligible nonspecific binding, ensuring high-contrast visualization of actin filaments, even in complex biological environments.
Phalloidin iFluor® 660 is optimized for high-resolution imaging and quantitative analysis of F-actin in diverse applications, including fixed tissue sections, cultured cells, and in vitro actin polymerization assays. Its compatibility with multiplex fluorescence imaging allows co-labeling with fluorescent proteins, quantum dots, iFluor® derivatives, and antibody-based detection systems.
Example protocol
AT A GLANCE
Prepare samples in microplate wells.
Remove liquid from samples in the plate.
Add Phalloidin-iFluor® 660 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 Cy5 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 STOCK SOLUTIONS
Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles
Add 30 µL of DMSO into the powder and mix well.
PREPARATION OF WORKING SOLUTION
Add 1 µL of Phalloidin-iFluor® 660 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 Phalloidin-iFluor® 660 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 (15) | |
Product family
| Name | Excitation (nm) | Emission (nm) | Extinction coefficient (cm -1 M -1) | Quantum yield | Correction Factor (260 nm) | Correction Factor (280 nm) |
| Phalloidin-iFluor® 350 Conjugate | 345 | 450 | 200001 | 0.951 | 0.83 | 0.23 |
| Phalloidin-iFluor® 405 Conjugate | 403 | 427 | 370001 | 0.911 | 0.48 | 0.77 |
| Phalloidin-iFluor® 488 Conjugate | 491 | 516 | 750001 | 0.91 | 0.21 | 0.11 |
| Phalloidin-iFluor® 514 Conjugate | 511 | 527 | 750001 | 0.831 | 0.265 | 0.116 |
| Phalloidin-iFluor® 532 Conjugate | 537 | 560 | 900001 | 0.681 | 0.26 | 0.16 |
| Phalloidin-iFluor® 555 Conjugate | 557 | 570 | 1000001 | 0.641 | 0.23 | 0.14 |
| Phalloidin-iFluor® 594 Conjugate | 587 | 603 | 2000001 | 0.531 | 0.05 | 0.04 |
| Phalloidin-iFluor® 633 Conjugate | 640 | 654 | 2500001 | 0.291 | 0.062 | 0.044 |
| Phalloidin-iFluor® 647 Conjugate | 656 | 670 | 2500001 | 0.251 | 0.03 | 0.03 |
Show More (4) | ||||||
Safety data sheet