Phalloidin Labeling Probes
Invitrogen™

Phalloidin Labeling Probes

Achieve precise and reliable F-actin staining with fluorescent and biotinylated phalloidins. Phalloidin conjugates are widely used in imaging applications to selectively label F-actin in a variety of sample types including fixed and permeabilized cells, tissue sections, and cell-free experiments.
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Catalog NumberColorExcitation Wavelength RangeDye Type
A22285Near-infrared663/690 nmAlexa Fluor™ 660
A22281Blue346/442 nmAlexa Fluor™ 350
A30104Violet405/450 nmAlexa Fluor™ Plus 405
A12379Green495/518 nmAlexa Fluor™ 488
O7466Green496/520 nmOregon Green™ 488
F432Green496/516 nmFITC (Fluorescein)
A22282Yellow531/554 nmAlexa Fluor™ 532
R415Red-orange540/565 nmTRITC
A22283Orange556/570 nmAlexa Fluor™ 546
A34055Orange555/565 nmAlexa Fluor™ 555
A30106Orange555/565 nmAlexa Fluor Plus 555
B3475Red558/569 nmBODIPY™
A12380Orange-red578/600 nmAlexa Fluor™ 568
A12381Red581/609 nmAlexa Fluor™ 594
T7471Red591/608 nmTexas Red™
A22284Far-red632/647 nmAlexa Fluor™ 633
A34054Far-red633/647 nmAlexa Fluor™ 635
A22287Far-red650/668 nmAlexa Fluor™ 647
A30107Far-red650/668 nmAlexa Fluor Plus 647
A22286Near-infrared679/702 nmAlexa Fluor™ 680
A30105Near-infrared758/784 nmAlexa Fluor™ Plus 750
B7474NoneNoneBiotin-XX
P3457NoneNonePhalloidin (unlabeled)
Catalog number A22285
Price (MXN)
-
Color:
Near-infrared
Excitation Wavelength Range:
663/690 nm
Dye Type:
Alexa Fluor™ 660
Fluorescent and biotinylated phalloidins are water soluble and bind to filamentous actin (F-actin) with nanomolar affinity, making them convenient probes for labeling, identifying, and quantifying F-actin in cryopreserved tissue sections, fixed and permeabilized cells, and cell-free experiments. Phalloidin conjugates bind similarly to actin from various species, including plants and animals, enabling staining of the cytoskeleton in a wide range of samples.

A variety of phalloidin conjugates for filamentous (F-actin) staining are available, including fluorescent Alexa Fluor and Alexa Fluor Plus phalloidins, along with phalloidins conjugated to classic fluorescent dyes such as BODIPY, fluorescein, and rhodamine. Phalloidin staining is spectrally compatible with other fluorescent stains used in cellular analyses such as GFP/RFP, Qdot nanocrystals, and other Alexa Fluor conjugates and antibodies. Biotin‐XX Phalloidin can be used to visualize actin filaments via fluorescent streptavidin tags or standard enzyme-mediated avidin/streptavidin techniques such as in electron microscopy. Unlabeled phalloidin is available for use as a control in blocking F‐actin staining or in promoting polymerization.

Phalloidin conjugates bind to both large and small actin filaments with similar affinity in a 1:1 stoichiometry between phallotoxin and actin subunits. They do not bind G-actin monomers.

Alexa Fluor and Alexa Fluor Plus phalloidin conjugates for F-actin staining

Fluorescent Alexa Fluor dye conjugates of phalloidin are popular F-actin stains, offering color choices across the full spectral range. These phalloidin conjugates provide researchers with fluorescent probes that are superior in brightness and photostability compared to other spectrally similar conjugates.

Alexa Fluor Plus Phalloidin conjugates retain the same specificity for actin but offer 3-5 times greater sensitivity and brightness compared to the corresponding Alexa Fluor Phalloidin conjugate. This increased brightness is beneficial for challenging F-actin imaging, such as the super‐resolution microscopy methods SIM and STORM, and for reliable staining of actin stress fibers.

Features of phalloidin probes

  • High specificity—binds selectively to F-actin, which allows for precise labeling of actin filaments in fixed cells and cryopreserved tissues
  • Strong affinity—nanomolar binding affinity for F-actin, which ensures stable and reliable actin staining
  • Extensive fluorescent conjugate options—over twenty conjugated varieties of phalloidin
  • Compatibility with fixed samples—typically used with fixed cells and tissues, making them suitable for actin staining in detailed structural studies, immunofluorescence staining, and IHC applications
  • Multiplexing capability—the wide availability of phalloidin conjugates enables their use in combination with other fluorescent probes and antibodies for multiplex imaging. Biotinylated phalloidin can be made use of in downstream streptavidin steps.
  • Quantitative analysis—can be used for quantitative analysis of F-actin distribution and density within cells, aiding in the study of cytoskeletal dynamics. The unlabeled phalloidin can be titrated as a control.
  • Ease of use—staining is straightforward and quick
  • Excellent stability—exhibit good photostability, which is essential for prolonged imaging sessions and time-lapse studies
  • Wide applicability—used for a range of applications, including studying cell morphology, motility, and the effects of drugs on the actin cytoskeleton
For Research Use Only. Not for use in diagnostic procedures.
Specifications
ColorNear-infrared
Dye TypeAlexa Fluor™ 660
Excitation Wavelength Range663/690 nm
For Use With (Equipment)Fluorescence Microscope, Flow Cytometer, Confocal Microscope, Compatible Cy5/Cy5.5 filter set
Product LineAlexa Fluor
Quantity300 Units
Shipping ConditionRoom Temperature
Label TypeAlexa Fluor Dyes
Product TypePhalloidin
Sub Cellular LocalizationActin, Cytoskeleton
Unit SizeEach
Contents & Storage
Store in freezer -5°C to -30°C and protect from light.

Frequently asked questions (FAQs)

Do you offer alternative package sizes for phalloidin or ReadyProbes reagents?

Alternative package sizes may be available through a custom request process. Contact Thermo Fisher Scientific customer support or your sales representative to discuss available options for the specific reagent and quantity needed.

Can I use conjugated phalloidin label probes on organoids?

Yes. Conjugated phalloidin labeling probes can be used with organoids when F-actin is accessible after fixation and permeabilization. Because organoids are thicker than monolayer cell cultures, protocol optimization may be needed, including permeabilization conditions, probe incubation time, and probe concentration.

Are conjugated phalloidin reagents antibodies?

No, Phalloidin is not an antibody. It is a bicyclic peptide toxin originally isolated from the Amanita phalloides mushroom and is used to bind and label F-actin in fixed and permeabilized samples.

Can I use methanol instead of DMSO to reconstitute the conjugated phalloidins from the lyophilized solids?

Yes, methanol can be used to reconstitute fluorescent phalloidin, but it creates a less concentrated stock solution than DMSO. Dissolve the vial contents in 1.5 mL of methanol to yield a 40X stock solution at a concentration of 200 assays/mL, which is equivalent to approximately 6.6 µM. One unit/assay of fluorescent phalloidins is equivalent to 5 µL of the methanolic stock solution. Methanol may be used as a stock-solution solvent, but methanol-containing fixatives are not recommended because they can disrupt F-actin.

How do I reconstitute fluorescent conjugated phalloidin probes to make a stock solution?

For fluorescent phalloidin label probes, prepare a DMSO stock solution by dissolving the vial contents in 150 µL anhydrous DMSO to make a 400X stock, with a concentration of approximately 66 µM or 2,000 assays per mL. Store between -25°C and -5°C, protected from light. One unit/assay of fluorescent phalloidins is equivalent to 0.5 µL of the DMSO stock solution.

Citations & References (15)

Citations & References
Abstract
Myosin Va maneuvers through actin intersections and diffuses along microtubules.
Authors:Ali MY, Krementsova EB, Kennedy GG, Mahaffy R, Pollard TD, Trybus KM, Warshaw DM
Journal:Proc Natl Acad Sci U S A
PubMed ID:17360524
'Certain types of intracellular organelle transport to the cell periphery are thought to involve long-range movement on microtubules by kinesin with subsequent handoff to vertebrate myosin Va (myoVa) for local delivery on actin tracks. This process may involve direct interactions between these two processive motors. Here we demonstrate using single ... More
Sidekicks: synaptic adhesion molecules that promote lamina-specific connectivity in the retina.
Authors:Yamagata M, Weiner JA, Sanes JR
Journal:Cell
PubMed ID:12230981
A major determinant of specific connectivity in the central nervous system is that synapses made by distinct afferent populations are restricted to particular laminae in their target area. We identify Sidekick (Sdk)-1 and -2, homologous transmembrane immunoglobulin superfamily molecules that mediate homophilic adhesion in vitro and direct laminar targeting of ... More
Two distinct distributions of F-actin are present in the hyphal apex of the oomycete Achlya bisexualis.
Authors:Yu YP, Jackson SL, Garrill A
Journal:Plant Cell Physiol
PubMed ID:15047875
We show that two distinct distributions of F-actin are present in the hyphal apex of the oomycete Achlya bisexualis, that have been chemically fixed with a combination of methylglyoxal and formaldehyde and stained with Alexa phalloidin. In approximately one half of the hyphae examined, an F-actin depleted zone within the ... More
A role for PKC-epsilon in Fc gammaR-mediated phagocytosis by RAW 264.7 cells.
Authors:Larsen EC, Ueyama T, Brannock PM, Shirai Y, Saito N, Larsson C, Loegering D, Weber PB, Lennartz MR
Journal:J Cell Biol
PubMed ID:12499353
Protein kinase C (PKC) plays a prominent role in immune signaling, and the paradigms for isoform selective signaling are beginning to be elucidated. Real-time microscopy was combined with molecular and biochemical approaches to demonstrate a role for PKC- epsilon in Fc gamma receptor (Fc gammaR)-dependent phagocytosis. RAW 264.7 macrophages were ... More
The HIV-1 pathogenicity factor Nef interferes with maturation of stimulatory T-lymphocyte contacts by modulation of N-Wasp activity.
Authors:Haller C, Rauch S, Michel N, Hannemann S, Lehmann MJ, Keppler OT, Fackler OT
Journal:J Biol Chem
PubMed ID:16687395
The Nef protein is a key determinant of human immunodeficiency virus (HIV) pathogenicity that, among other activities, sensitizes T-lymphocytes for optimal virus production. The initial events by which Nef modulates the T-cell receptor (TCR) cascade are poorly understood. TCR engagement triggers actin rearrangements that control receptor clustering for signal initiation ... More