Phalloidin Conjugates

iFluor® 405

This blue fluorescent phalloidin conjugate (equivalent to Alexa Fluor® 405-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™ 405 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 DAPI filter

Important Warm the vial to room temperature and centrifuge briefly before opening.

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™ 405 Conjugate working solution

Add 1 µL of Phalloidin-iFluor™ 405 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™ 405 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 DAPI filter set.

Spectrum

Open in Advanced Spectrum Viewer

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
Phalloidin-California Red Conjugate	California Red
Phalloidin-Tetramethylrhodamine Conjugate	TRITC
Phalloidin-iFluor® 700 Conjugate	iFluor 700
Phalloidin-AMCA Conjugate	AMCA
Phalloidin-Biotin Conjugate	Biotin
Phalloidin-iFluor® 790 Conjugate	iFluor 790
XFD594 Phalloidin XFD594 Same Structure to Alexa Fluor™ 594	Alexa Fluor 594
Phalloidin-iFluor® 750 Conjugate	iFluor 750
Phalloidin-iFluor® 680 Conjugate	iFluor 680
Phalloidin-Fluorescein Conjugate	Fluorescein
XFD350 Phalloidin XFD350 Same Structure to Alexa Fluor™ 350	Alexa Fluor 350
XFD488 Phalloidin XFD488 Same Structure to Alexa Fluor™ 488	Alexa Fluor 488
XFD647 Phalloidin equivalent to Alexa Fluor® 647 phalloidin	Alexa Fluor 647
Phalloidin CAS#: 17466-45-4	Unlabeled
Phalloidin-iFluor® 660 Conjugate	iFluor 660
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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® 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
Phalloidin-iFluor® 680 Conjugate	684	701	220000¹	0.23¹	0.097	0.094
Phalloidin-iFluor® 700 Conjugate	690	713	220000¹	0.23¹	0.09	0.04
Phalloidin-iFluor® 750 Conjugate	757	779	275000¹	0.12¹	0.044	0.039
Phalloidin-iFluor® 790 Conjugate	787	812	250000¹	0.13¹	0.1	0.09
Phalloidin-iFluor® 660 Conjugate	663	678	250000¹	0.26¹	0.07	0.08
iFluor® 405-streptavidin conjugate	403	427	37000¹	0.91¹	0.48	0.77
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Citations

View all 56 citations: Citation Explorer

Pyroptotic cell corpses are crowned with F-actin-rich filopodia that engage CLEC9A signaling in incoming dendritic cells
Authors: Holley, Caroline L and Monteleone, Mercedes and Fisch, Daniel and Libert, Alexandre ES and Ju, Robert J and Choi, Joon H and Condon, Nicholas D and Emming, Stefan and Crawford, Joanna and Lawrence, Grace MEP and others,
Journal: Nature Immunology (2024): 1--11

Hippo signaling pathway regulates Ebola virus transcription and egress
Authors: Liang, Jingjing and Djurkovic, Marija A and Leavitt, Carson G and Shtanko, Olena and Harty, Ronald N
Journal: Nature Communications (2024): 6953

A novel delta opioid receptor specific peptide reduces craving in an animal model of cocaine seeking
Authors: Itzhak, Pnina Shirel and Yael, Hevroni and Matsree, Erez and Pee'r-Nissan, Hilla and Lancman, Shira Ofer and Barnea, R and Luboshits, G and Motiei, Menachem and Betzer, Oshra and Gispan, Iris and others,
Journal: Addiction Neuroscience (2024): 100159

Osteoblast cell behavior on polyetheretherketone dental implant surfaces treated with different grit size aluminum oxide particles: An in vitro analysis
Authors: Gaikwad, Amit and Parizi, Marjan Kheirmand and Winkel, Andreas and Stiesch, Meike
Journal: The Journal of Prosthetic Dentistry (2024)

mRNA-Laden Lipid-Nanoparticle-Enabled in Situ CAR-Macrophage Engineering for the Eradication of Multidrug-Resistant Bacteria in a Sepsis Mouse Model
Authors: Tang, Chunwei and Jing, Weiqiang and Han, Kun and Yang, Zhenmei and Zhang, Shengchang and Liu, Miaoyan and Zhang, Jing and Zhao, Xiaotian and Liu, Ying and Shi, Chongdeng and others,
Journal: ACS nano (2024)

References

View all 127 references: Citation Explorer

Improved penile histology by phalloidin stain: circular and longitudinal cavernous smooth muscles, dual-endothelium arteries, and erectile dysfunction-associated changes
Authors: Lin G, Qiu X, F and el TM, Albersen M, Wang Z, Lue TF, Lin CS.
Journal: Urology (2011): 970 e1

Phalloidin perturbs the interaction of human non-muscle myosin isoforms 2A and 2C1 with F-actin
Authors: Diensthuber RP, Muller M, Heissler SM, Taft MH, Chizhov I, Manstein DJ.
Journal: FEBS Lett (2011): 767

pH-(low)-insertion-peptide (pHLIP) translocation of membrane impermeable phalloidin toxin inhibits cancer cell proliferation
Authors: An M, Wijesinghe D, Andreev OA, Reshetnyak YK, Engelman DM.
Journal: Proc Natl Acad Sci U S A (2010): 20246

Labeling cytoskeletal F-actin with rhodamine phalloidin or fluorescein phalloidin for imaging
Authors: Chazotte B., undefined
Journal: Cold Spring Harb Protoc (2010): pdb prot4947

Effect of Phalloidin on Filaments Polymerized from Heart Muscle Adp-Actin Monomers
Authors: Vig A, Dudas R, Kupi T, Orban J, Hild G, Lorinczy D, Nyitrai M.
Journal: J Therm Anal Calorim (2009): 721

Page updated on October 9, 2024

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Catalog Number	23100231012310223103231102311123115231162311723119231222312523126231272312823129231302313123140231502315323158231595301
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Physical properties

Molecular weight	~1100
Solvent	DMSO

Spectral properties

Correction Factor (260 nm)	0.48
Correction Factor (280 nm)	0.77
Extinction coefficient (cm ^-1 M ^-1)	37000¹
Excitation (nm)	403
Emission (nm)	427
Quantum yield	0.91¹

Storage, safety and handling

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