Phalloidin-iFluor® 647 Conjugate
Ordering information
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Additional ordering information
Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
Quotation | Request |
International | See distributors |
Shipping | Standard overnight for United States, inquire for international |
Physical properties
Molecular weight | 1408.65 |
Solvent | DMSO |
Spectral properties
Correction Factor (260 nm) | 0.03 |
Correction Factor (280 nm) | 0.03 |
Correction Factor (656 nm) | 0.0793 |
Extinction coefficient (cm -1 M -1) | 2500001 |
Excitation (nm) | 656 |
Emission (nm) | 670 |
Quantum yield | 0.251 |
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 | ![]() ![]() |
See also: Antibodies and Proteomics, Antibody and Protein Labeling, Bioconjugation, Cell Structures and Organelles, Actin, iFluor® Dyes and Kits, Phalloidin Conjugates
Molecular weight 1408.65 | Correction Factor (260 nm) 0.03 | Correction Factor (280 nm) 0.03 | Correction Factor (656 nm) 0.0793 | Extinction coefficient (cm -1 M -1) 2500001 | Excitation (nm) 656 | Emission (nm) 670 | Quantum yield 0.251 |
This deep red fluorescent phalloidin conjugate (equivalent to Alexa Fluor® 647-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™ 647 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
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 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.
Phalloidin-iFluor™ 647 Conjugate stock solution
Add 30 µL of DMSO into the powder and mix well.PREPARATION OF WORKING SOLUTION
Phalloidin-iFluor™ 647 Conjugate working solution
Add 1 µL of Phalloidin-iFluor™ 647 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™ 647 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.
Calculators
Common stock solution preparation
Table 1. Volume of DMSO needed to reconstitute specific mass of Phalloidin-iFluor® 647 Conjugate to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.
0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
1 mM | 70.99 µL | 354.95 µL | 709.9 µL | 3.549 mL | 7.099 mL |
5 mM | 14.198 µL | 70.99 µL | 141.98 µL | 709.9 µL | 1.42 mL |
10 mM | 7.099 µL | 35.495 µL | 70.99 µL | 354.95 µL | 709.9 µL |
Molarity calculator
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Spectrum
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Spectral properties
Correction Factor (260 nm) | 0.03 |
Correction Factor (280 nm) | 0.03 |
Correction Factor (656 nm) | 0.0793 |
Extinction coefficient (cm -1 M -1) | 2500001 |
Excitation (nm) | 656 |
Emission (nm) | 670 |
Quantum yield | 0.251 |
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 | 588 | 604 | 1800001 | 0.531 | 0.05 | 0.04 |
Phalloidin-iFluor® 633 Conjugate | 640 | 654 | 2500001 | 0.291 | 0.062 | 0.044 |
Phalloidin-iFluor® 680 Conjugate | 684 | 701 | 2200001 | 0.231 | 0.097 | 0.094 |
Show More (5) |
Images

Figure 1. Fluorescence images of HeLa cells stained with Phalloidin-iFluor® 647 Conjugate using fluorescence microscope with a Cy5 filter set (Red). Live cells were first stained with mitochondria dye MitoLite™ Green. After fixation in 4% formaldehyde, cells were labeled with Phalloidin-iFluor® 647 and counterstained with Nuclear Blue™ DCS1 (Cat#17548, Blue).

Figure 2. VG and IF images of the ORL. The periosteal (row A), intramuscular (row B), preorbicularis (row C) and dermal (row D) regions of the ORL were observed by VG (column 1) and IF (columns 2–5). There were immunopositive reactions for elastin (column 2, blue), collagen type I (column 3, green) and actin (column 4, red). The column 5 shows a merged image of the images in columns 2–4 image. Asterisks indicate confluence of the perimysium into the ORL fibres. Source: Three-dimensional structure of the orbicularis retaining ligament: an anatomical study using micro-computed tomography by Jehoon O et al., Scientific Reports, Nov. 2018.

Figure 3. Overall structure of the orbicularis retaining ligament (ORL). (a) Three-dimensional (3D) morphology reconstructed from micro-computed tomography (mCT) image sections. (b) Modified Verhoeff Van Gieson staining (VG) image. (c) A merged immunofluorescence (IF) image (elastin, blue; collagen type I, green; actin, red). Arrowheads indicate a direct fibre from the periosteum (P) to the dermis (D). OOc, orbicularis oculi muscle. S, sagittal; M, medial; A, anterior. Source: Three-dimensional structure of the orbicularis retaining ligament: an anatomical study using micro-computed tomography by Jehoon O et al., Scientific Reports, Nov. 2018.

Figure 5. Fluorescence images of HeLa cells stained with Phalloidin-iFluor® 647 Conjugate using fluorescence microscope with a Cy5 filter set (Red). HeLa cells were fixed with 4% formaldehyde followed by incubation with 1 ug/mL mouse tubulin antibody. Cells were stained with 10 ug/mL of GxM IgG- iFluor 488 conjugates. Cells were stained with Phalloidin-iFluor® 647 conjugate following product protocol and incubated with 2 uM DAPI for 5 min before imaging.

Figure 6. Co-localization of caveolin-1 and actin in post-conditioned hearts treated with latrunculin A. Representative confocal laser microscopy of the sections of frozen heart tissue after double-immunofluorescence staining of caveolin-1 (green) and actin (red). Cell nuclei were counterstained with DAPI. Source: Actin-Cytoskeleton Drives Caveolae Signaling to Mitochondria during Postconditioning by Correa et. al., Cells. Feb. 2023.

Figure 7. Cytokines induce intracellular component and cytoskeleton changes in RA synoviocytes. Holographic and tomographic microscopy images and phalloidin staining. Phalloidin-iFluor 647 was used for staining. Source: Effects of pro-inflammatory cytokines and cell interactions on cell area and cytoskeleton of rheumatoid arthritis synoviocytes and immune cells by Filali et. al., European Journal of Cell Biology. March 2023
Citations
View all 86 citations: Citation Explorer
Generation and characterization of human oligodendroglia as cellular model for Multiple System Atrophy
Authors: Wihan, Jeanette
Journal: (2023)
Authors: Wihan, Jeanette
Journal: (2023)
Effects of pro-inflammatory cytokines and cell interactions on cell area and cytoskeleton of rheumatoid arthritis synoviocytes and immune cells
Authors: Filali, Samira and Noack, M{\'e}lissa and G{\'e}lo{\"e}n, Alain and Pirot, Fabrice and Miossec, Pierre
Journal: European Journal of Cell Biology (2023): 151303
Authors: Filali, Samira and Noack, M{\'e}lissa and G{\'e}lo{\"e}n, Alain and Pirot, Fabrice and Miossec, Pierre
Journal: European Journal of Cell Biology (2023): 151303
Egg white improves the biological properties of an alginate-methylcellulose bioink for 3D bioprinting of volumetric bone constructs
Authors: Liu, Suihong and Kilian, David and Ahlfeld, Tilman and Hu, Qingxi and Gelinsky, Michael
Journal: Biofabrication (2023)
Authors: Liu, Suihong and Kilian, David and Ahlfeld, Tilman and Hu, Qingxi and Gelinsky, Michael
Journal: Biofabrication (2023)
Human iPSC-Derived Proinflammatory Macrophages cause Insulin Resistance in an Isogenic White Adipose Tissue Microphysiological System
Authors: Qi, Lin and Matsuo, Koji and Pereira, Ashley and Lee, Yue Tung and Zhong, Fenmiao and He, Yuchen and Zushin, Peter-James H and Gr{\"o}ger, Marko and Sharma, Aditi and Willenbring, Holger and others,
Journal: Small (2023): 2203725
Authors: Qi, Lin and Matsuo, Koji and Pereira, Ashley and Lee, Yue Tung and Zhong, Fenmiao and He, Yuchen and Zushin, Peter-James H and Gr{\"o}ger, Marko and Sharma, Aditi and Willenbring, Holger and others,
Journal: Small (2023): 2203725
Targeting host SUR2 can reduce severity of Helicobacter pylori associated gastritis Targeting SUR2 to reduce H. pylori associated gastritis
Authors: Sarkar, Sohinee and Talesh, Ghazal Alipour and Menheniott, Trevelyan R and Sutton, Philip
Journal: Gastro Hep Advances (2023)
Authors: Sarkar, Sohinee and Talesh, Ghazal Alipour and Menheniott, Trevelyan R and Sutton, Philip
Journal: Gastro Hep Advances (2023)
Inhibition of Exchange Proteins Directly Activated by cAMP (EPAC) as a Strategy for Broad-Spectrum Antiviral Development
Authors: Boulton, Stephen and Crupi, Mathieu JF and Singh, Siddharth and Carter-Timofte, Madalina E and Azad, Taha and Organ, Bailey C and He, Xiaohong and Gill, Rida and Neault, Serge and Jamieson, Taylor and others,
Journal: Journal of Biological Chemistry (2023): 104749
Authors: Boulton, Stephen and Crupi, Mathieu JF and Singh, Siddharth and Carter-Timofte, Madalina E and Azad, Taha and Organ, Bailey C and He, Xiaohong and Gill, Rida and Neault, Serge and Jamieson, Taylor and others,
Journal: Journal of Biological Chemistry (2023): 104749
The Enzyme 15-Hydroxyprostaglandin Dehydrogenase Inhibits a Shift to the Mesenchymal Pattern of Trophoblasts and Decidual Stromal Cells Accompanied by Prostaglandin Transporter in Preeclampsia
Authors: Pang, Huiyuan and Lei, Di and Chen, Tingting and Liu, Yujie and Fan, Cuifang
Journal: International Journal of Molecular Sciences (2023): 5111
Authors: Pang, Huiyuan and Lei, Di and Chen, Tingting and Liu, Yujie and Fan, Cuifang
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Vacuole Membrane Protein 1 (VMP1) Restricts NLRP3 Inflammasome Activation by Modulating SERCA Activity and Autophagy
Authors: Zack, Stephanie R and Nikolaienko, Roman and Cook, Ben and Melki, Ronald and Zima, Aleksey V and Campbell, Edward M
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Disrupting the $\alpha$-synuclein-ESCRT interaction with a peptide inhibitor mitigates neurodegeneration in preclinical models of Parkinson’s disease
Authors: Nim, Satra and O’Hara, Darren M and Corbi-Verge, Carles and Perez-Riba, Albert and Fujisawa, Kazuko and Kapadia, Minesh and Chau, Hien and Albanese, Federica and Pawar, Grishma and De Snoo, Mitchell L and others,
Journal: Nature Communications (2023): 2150
Authors: Nim, Satra and O’Hara, Darren M and Corbi-Verge, Carles and Perez-Riba, Albert and Fujisawa, Kazuko and Kapadia, Minesh and Chau, Hien and Albanese, Federica and Pawar, Grishma and De Snoo, Mitchell L and others,
Journal: Nature Communications (2023): 2150
Transient receptor potential channel vanilloid 1 (TRPV1) contributes to facial mechanical hypersensitivity in a mouse model of atopic asthma
Authors: Cao, Ailin and Gao, Weiqi and Sawada, Takeshi and Yoshimoto, Reiko U and Aijima, Reona and Ohsaki, Yasuyoshi and Kido, Mizuho A
Journal: Laboratory Investigation (2023): 100149
Authors: Cao, Ailin and Gao, Weiqi and Sawada, Takeshi and Yoshimoto, Reiko U and Aijima, Reona and Ohsaki, Yasuyoshi and Kido, Mizuho A
Journal: Laboratory Investigation (2023): 100149
References
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