Description
Key facts
Applications:Fluorescence Microscopy, ICC/IFSee reactivity dataSee the reactivity data table below for information on validated species and application combinations.,
Target:ACTBSee target data,
Storage buffer:Constituents: 100% Fluorescent-Phalloidin Conjugate
Product details:
How the assay works
Phalloidin-iFluor 647 Reagent (equivalent to Alexa Fluor® 647-labeled phalloidin) selectively binds to F-actins. Phalloidin-iFluor 647 can be detected with a fluorescent microscope at Ex/Em = 650/665 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 647 Reagent Protocol Summary
- Prepare samples in microplate wells
- Remove liquid from samples in the plate
- Add Phalloidin-iFluor™ 647 (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
How other researchers are using
Phalloidin-iFluor 647 Reagent has been used in a variety of sample type including:
- Human Induced Pluripotent Stem Cells 1
- Human BrCa cell line MCF7 and T47D 2
- HEK293T cells 3
References:
1 - Doni D et al. 2024; 2 - Qiu Y et al. 2023; 3 - Wang X et al. 2023.
Related and recommended products
Review other popular phalloidin dye conjugates, including Phalloidin-iFluor 488 (ab176753), Phalloidin-iFluor 555 (ab176756) and Rhodamine Phalloidin (ab235138), or read the
phalloidin staining protocol
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
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.
Biological function summary
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.
Pathways
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.
