Phalloidin-iFluor® 594 Conjugate
Ordering information
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Additional ordering information
Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
International | See distributors |
Bulk request | Inquire |
Custom size | Inquire |
Shipping | Standard overnight for United States, inquire for international |
Physical properties
Molecular weight | ~1600 |
Solvent | DMSO |
Spectral properties
Absorbance (nm) | 587 |
Correction Factor (260 nm) | 0.05 |
Correction Factor (280 nm) | 0.04 |
Extinction coefficient (cm -1 M -1) | 1800001 |
Excitation (nm) | 588 |
Emission (nm) | 604 |
Quantum yield | 0.531 |
Storage, safety and handling
Certificate of Origin | Download PDF |
H-phrase | H301, H311, H331 |
Hazard symbol | T |
Intended use | Research Use Only (RUO) |
R-phrase | R23, R24, R25 |
Storage | Freeze (< -15 °C); Minimize light exposure |
UNSPSC | 12352200 |
Overview | ![]() ![]() |
Molecular weight ~1600 | Absorbance (nm) 587 | Correction Factor (260 nm) 0.05 | Correction Factor (280 nm) 0.04 | Extinction coefficient (cm -1 M -1) 1800001 | Excitation (nm) 588 | Emission (nm) 604 | Quantum yield 0.531 |
This red fluorescent phalloidin conjugate (equivalent to Alexa Fluor® 594-labeled phalloidin) selectively binds to F-actins. 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. The property of phalloidin is a useful tool for investigating the distribution of F-actin in cells by labeling phalloidin with fluorescent analogs and using them to stain actin filaments for light microscopy. 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. AAT Bioquest offers a variety of fluorescent phalloidin derivatives with different colors for multicolor imaging applications.
Example protocol
AT A GLANCE
Protocol Summary
- Prepare samples in microplate wells
- Remove liquid from samples in the plate
- Add Phalloidin-iFluor™ 594 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 TRITC or Texas Red filter
Storage and Handling Conditions
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
Phalloidin-iFluor™ 594 Conjugate working solution
Add 1 µL of Phalloidin-iFluor™ 594 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
Stain the cells
- 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™ 594 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 TRITC or Texas Red filter set.
Spectrum
Open in Advanced Spectrum Viewer


Spectral properties
Absorbance (nm) | 587 |
Correction Factor (260 nm) | 0.05 |
Correction Factor (280 nm) | 0.04 |
Extinction coefficient (cm -1 M -1) | 1800001 |
Excitation (nm) | 588 |
Emission (nm) | 604 |
Quantum yield | 0.531 |
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® 633 Conjugate | 640 | 654 | 2500001 | 0.291 | 0.062 | 0.044 |
Phalloidin-iFluor® 647 Conjugate | 656 | 670 | 2500001 | 0.251 | 0.03 | 0.03 |
Phalloidin-iFluor® 680 Conjugate | 684 | 701 | 2200001 | 0.231 | 0.097 | 0.094 |
Show More (4) |
Images

Figure 1. Fluorescence image of HeLa cells fixed with 4% formaldehyde then stained with Cell Navigator® F-Actin Labeling Kit *Red Fluorescence* in a Costar black 96-well plate. Cells were labeled with Phalloidin-iFluor® 594 (Cat#23122, Red) and nuclei stain DAPI (Cat#17507, Blue), respectively. Cell endoplasmic reticulum (ER) was stained with ER Green™ (Cat#22635, Green) before fixation.

Figure 2. Distribution of MCs in the membranous cochlea. The MCs were scored in each cochlear preparation, and the results are presented as a total number of MCs (non-degranulated and degranulated) per area ((a) n = 7; (b) n = 9). (a) Violin plot demonstrating MCs distribution in the explant areas containing either spiral limbus with OC or the lateral wall; (b) Violin plot demonstrating MCs distribution in the apical, medial, and basal parts of the cochlea; the marker shows the mean. (c–f) Representative micrographs showing the cochlear MCs in the medial part of the cochlea. The explants were stained with phalloidin-iFluor 594, avidin–Alexa Fluor ™ 488, and DAPI. The arrows point to MCs. (f) contains a digital enlargement of the MC image form (e). The data were derived from two independent experiments, and the differences were calculated for means. “ns” indicates not significant (p > 0.05); * p < 0.05; *** p < 0.001. ((a) Kruskal–Wallis test; (b) Mann–Whitney test). Source: Degranulation of Murine Resident Cochlear Mast Cells: A Possible Factor Contributing to Cisplatin-Induced Ototoxicity and Neurotoxicity by Karayay et. al., Int. J. Mol. Sci. Feb. 2023

Figure 3. Cromolyn used at high concentrations decreases the numbers of IHCs and OHCs. (a) The intact hair cells were scored along the length of 100 µm of the cochlear explant (spiral limbus containing OC), and the percentages of intact IHCs (red circles) and OHCs (green squares) were determined and plotted on the y-axis. The control explants (n = 4) were cultured for 24 h in a tissue culture medium. The treatment groups (n = 4 for each treatment) were cultured for 24 h with cromolyn at the following concentrations: 5 µM, 10 µM, 25 µM, 50 µM, 100 µM, and 200 µM. (b,c) Representative micrograph showing intact (b) and damaged (c) hair cells. Arrows point out intact HCs (white) and damaged HCs (green). Scale bar represents 10 µM. (d–i) Representative micrograph showing the HCs after 24 h of exposure to 5 µM, 10 µM, 25 µM, 50 µM, 100 µM, and 200 µM cromolyn. The cochlea explants were stained with phalloidin-iFluor 594. The scale bar represents 10 µm. Four independent experiments were performed; the data are reported as mean ± SEM. * p < 0.05; ** p < 0.01 (two-way ANOVA with Dunnett multiple comparison test). Source: Degranulation of Murine Resident Cochlear Mast Cells: A Possible Factor Contributing to Cisplatin-Induced Ototoxicity and Neurotoxicity by Karayay et.al., Int. J. Mol. Sci. Feb. 2023
Citations
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Application notes
A Meta-Analysis of Common Calcium Indicators
A New Protein Crosslinking Method for Labeling and Modifying Antibodies
A Novel Fluorescent Probe for Imaging and Detecting Hydroxyl Radical in Living Cells
A Novel NO Wash Probeniceid-Free Calcium Assay for Functional Analysis of GPCR and Calcium Channel Targets
Abbreviation of Common Chemical Compounds Related to Peptides
A New Protein Crosslinking Method for Labeling and Modifying Antibodies
A Novel Fluorescent Probe for Imaging and Detecting Hydroxyl Radical in Living Cells
A Novel NO Wash Probeniceid-Free Calcium Assay for Functional Analysis of GPCR and Calcium Channel Targets
Abbreviation of Common Chemical Compounds Related to Peptides
FAQ
Are Cell Navigator® Cell Plasma Membrane Staining Kits suitable for cell culture medium samples?
Are there any alternatives to BrdU (Bromodeoxyuridine)?
Are there any alternatives to Cy5?
Are there any alternatives to indocyanine green (ICG)?
Are there any calcium indicators that don't require probenecid (PBC)?
Are there any alternatives to BrdU (Bromodeoxyuridine)?
Are there any alternatives to Cy5?
Are there any alternatives to indocyanine green (ICG)?
Are there any calcium indicators that don't require probenecid (PBC)?