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
A22283Orange556/570 nmAlexa Fluor™ 546
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
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
A22285Near-infrared663/690 nmAlexa Fluor™ 660
A22286Near-infrared679/702 nmAlexa Fluor™ 680
A30105Near-infrared758/784 nmAlexa Fluor™ Plus 750
B7474NoneNoneBiotin-XX
P3457NoneNonePhalloidin (unlabeled)
Catalog number A22283
Price (MXN)
-
Color:
Orange
Excitation Wavelength Range:
556/570 nm
Dye Type:
Alexa Fluor™ 546
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
ColorOrange
Dye TypeAlexa Fluor™ 546
Excitation Wavelength Range556/570 nm
For Use With (Equipment)Fluorescence Microscope, Flow Cytometer, Confocal Microscope, Compatible with TRITC/RFP 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 (33)

Citations & References
Abstract
DOCK2 is a Rac activator that regulates motility and polarity during neutrophil chemotaxis.
Authors:Kunisaki Y,Nishikimi A,Tanaka Y,Takii R,Noda M,Inayoshi A,Watanabe K,Sanematsu F,Sasazuki T,Sasaki T,Fukui Y
Journal:The Journal of cell biology
PubMed ID:16943182
Neutrophils are highly motile leukocytes, and they play important roles in the innate immune response to invading pathogens. Neutrophil chemotaxis requires Rac activation, yet the Rac activators functioning downstream of chemoattractant receptors remain to be determined. We show that DOCK2, which is a mammalian homologue of Caenorhabditis elegans CED-5 and ... More
Expression of the voltage-gated sodium channel NaV1.5 in the macrophage late endosome regulates endosomal acidification.
Authors:Carrithers MD, Dib-Hajj S, Carrithers LM, Tokmoulina G, Pypaert M, Jonas EA, Waxman SG,
Journal:J Immunol
PubMed ID:17548620
'Voltage-gated sodium channels expressed on the plasma membrane activate rapidly in response to changes in membrane potential in cells with excitable membranes such as muscle and neurons. Macrophages also require rapid signaling mechanisms as the first line of defense against invasion by microorganisms. In this study, our goal was to ... More
Time-lapse analysis and mathematical characterization elucidate novel mechanisms underlying muscle morphogenesis.
Authors:Snow CJ, Goody M, Kelly MW, Oster EC, Jones R, Khalil A, Henry CA,
Journal:PLoS Genet
PubMed ID:18833302
'Skeletal muscle morphogenesis transforms short muscle precursor cells into long, multinucleate myotubes that anchor to tendons via the myotendinous junction (MTJ). In vertebrates, a great deal is known about muscle specification as well as how somitic cells, as a cohort, generate the early myotome. However, the cellular mechanisms that generate ... More
Characterizing peptide-mediated DNA internalization in human cancer cells.
Authors:Wittrup A, Belting M,
Journal:Methods Mol Biol
PubMed ID:19085116
'Cell penetrating peptides (CPPs) are currently used to deliver various macromolecular cargos to intracellular sites of action both in vitro and in vivo on an experimental basis. During the last few years, even more evidence has accumulated indicating that the main route of entry for most CPPs is through endocytosis ... More
Zinquin identifies subcellular compartmentalization of zinc in cortical neurons. Relation to the trafficking of zinc and the mitochondrial compartment.
Authors:Colvin RA, Laskowski M, Fontaine CP,
Journal:Brain Res
PubMed ID:16581038
'Zinquin (Zn(2+) selective fluorophore), when used to visualize intracellular Zn(2+), typically shows brightly fluorescent perinuclear endosome-like structures, presumably identifying Zn(2+) containing organelles. In this study, zinquin identified numerous and widespread sites of Zn(2+) compartmentalization in primary cultures of embryonic rat cortical neurons. Nuclear fluorescence, however, was absent. We labeled neuronal ... More