Phalloidin-iFluor 555 reagent (ab176756) is used to label F-actin (actin filaments) with high affinity and low background. The readout is on any fluorescent microscope with Ex/Em = 556/574 nm.
- Can be used for fluorescent co-labelling
- Broad sample compatibility- including tissue sections, cell cultures
- Cited in over 80 publications
Select an associated product type
Actin is a highly conserved protein that polymerizes to produce filaments that form cross-linked networks in the cytoplasm of cells (PubMed:25255767, PubMed:29581253). Actin exists in both monomeric (G-actin) and polymeric (F-actin) forms, both forms playing key functions, such as cell motility and contraction (PubMed:29581253). In addition to their role in the cytoplasmic cytoskeleton, G- and F-actin also localize in the nucleus, and regulate gene transcription and motility and repair of damaged DNA (PubMed:29925947). Part of the ACTR1A/ACTB filament around which the dynactin complex is built. The dynactin multiprotein complex activates the molecular motor dynein for ultra-processive transport along microtubules (By similarity).
Beta-actin, ACTB
Phalloidin-iFluor 555 reagent (ab176756) is used to label F-actin (actin filaments) with high affinity and low background. The readout is on any fluorescent microscope with Ex/Em = 556/574 nm.
- Can be used for fluorescent co-labelling
- Broad sample compatibility- including tissue sections, cell cultures
- Cited in over 80 publications
How the assay works
Phalloidin-iFluor 555 Reagent (equivalent to Alexa Fluor® 555-labeled phalloidin) selectively binds to F-actins.It can be detected with a fluorescent microscope at Ex/Em = 556/574 nm. Phalloidin conjugates are convenient probes for labeling, identifying and quantifying animal or plant actin filaments in formaldehyde-fixed and permeabilized tissue sections, cell cultures or cell-free experiments. They can also be used in paraffin-embedded samples that have been de-paraffinized. Staining fixed cell or tissue samples with phalloidin conjugates is very simple; it requires a single 20-90 min incubation with the phalloidin, followed by 3 short wash steps. Phalloidin staining can be combined with antibody-based staining by adding the phalloidin conjugate during either the primary or secondary antibody incubation step.
When used in unfixed samples, phalloidin binding leads to a decrease in the disassociation rate of actin subunits from the ends of actin filaments, essentially stabilizing actin filaments through the prevention of filament depolymerisation.
Phalloidin-iFluor 555 Reagent Protocol Summary
- Prepare samples in microplate wells
- Remove liquid from samples in the plate
- Add Phalloidin-iFluor 555 (100 µL/well)
- Stain the cells at room temperature for 20 to 90 minutes
- Wash the cells
- Examine the specimen under microscope with TRITC filter
How other researchers are using
Phalloidin-iFluor 555 Reagent has been used in a variety of sample type including:
- Rabbit corneas 1
- Human Mesodermal Progenitor Cells 2
- Humanumbilical vein endothelial cell line 3
References:
1 - Song E et al. 2024; 2 - Stцberl N et al. 2023; 3 - Songjang W et al. 2023.
Related and recommended products
Review other popular phalloidin dye conjugates, including Phalloidin-iFluor 488 (Phalloidin-iFluor 488 Reagent ab176753), Phalloidin-iFluor 647 (Phalloidin-iFluor 647 Reagent ab176759) and Rhodamine Phalloidin (Rhodamine Phalloidin Reagent ab235138), or read the phalloidin staining protocol.
F-actin also known as filamentous actin is an essential structural protein found within the cytoskeleton of eukaryotic cells. Its alternate names include actin filaments or microfilaments. The protein consists of polymerized monomers of G-actin each with a molecular weight of roughly 42 kDa. F-actin is expressed abundantly in muscle cells and non-muscle cells alike providing structural support and facilitating cellular movements. Actin staining is a common method used in labs to visualize these dynamic structures often employing phalloidin staining a toxin that stabilizes actin filaments conjugated with fluorescent labels such as Phalloidin 594 Phalloidin 647 or Phalloidin 488 for imaging purposes.
The actin cytoskeleton plays integral roles in maintaining cell shape providing mechanical resistance against deformation and driving important cellular processes such as endocytosis cell division and motility. F-actin forms part of numerous protein complexes interacting with other proteins like myosin to facilitate muscle contraction and cellular transport. Within cells F-actin is dynamic readily polymerizing and depolymerizing in response to cellular signaling making it essential for cytoskeletal remodeling and cellular adaptability.
F-actin is central to various signaling cascades underlying processes like cell signaling and intracellular transport. Notably it participates in the Rho family GTPase pathway affecting cell cytoskeleton organization and motility. It also interacts with proteins like cofilin and profilin which regulate actin polymerization and treadmilling dynamics respectively. These interactions highlight F-actin's involvement in complex cellular pathway regulation processes essential for maintaining cellular homeostasis and adaptability.
Abnormal regulation or mutations in actin-related proteins can lead to conditions such as cancer and cardiomyopathies. For example during metastasis cancer cells exploit the dynamic nature of F-actin for enhanced migratory capacity. In cardiac muscle cells actin interacts with other proteins like tropomyosin and mutations in these genes can disrupt normal heart function leading to cardiomyopathies. As such F-actin not only represents a critical component of the cellular structure but also serves as a pivotal target for understanding disease mechanisms and potential therapeutic intervention points.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
Actin filaments staining in HeLa cells. Actin filaments (red) were stained with CytoPainter Phalloidin-iFluor 555 reagent (Phalloidin-iFluor 532 Reagent ab176755). Nuclei were stained with Nuclear Green DCS1 (Nuclear Green DCS1 ab138905).
Immunofluorescent images of Phalloidin staining at day 3 across various surface thicknesses. Red fluorescence indicates phalloidin staining for F-actin and blue fluorescence indicates DAPI staining for cell nuclei.
Excitation and emission of phalloidin-iFluor 555 reagent.
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com