Phalloidin-iFluor 405 Reagent
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(1 Review)
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(35 Publications)
Phalloidin-iFluor 405 Reagent ab176752 is one of a series of phalloidin conjugates that bind to actin filaments, also known as F-actin.
View Alternative Names
Beta-actin, ACTB
- ICC/IF
AbReview49547****
Immunocytochemistry/ Immunofluorescence - Phalloidin-iFluor 405 Reagent (AB176752)
CytoPainter Phalloidin-iFluor 405 Reagent was used to stain F-actin in Bovine Foetal Aeorta Endothelial (BFA) cells. This was tested on triton x-100 permeabilised and non-permeabilised cells, phalloidin was prepared at 1 in 1000, diluted in 1% BSA in PBS. Cells were stained for 20 minutes. Coverslips with cells were then prepared and used for confocal microscopy. Nucleus was also stained with a nuclear stain that is excited by 633 laser. ctin staining worked in both permeabilised and unpermeabilised cells, although fluorescence was dimmer when compared to phalloidin-488 or phalloidin-568 stain, however this could be overcome perhaps by leaving the product on the cells for longer. Fluoresence also bleached relatively quickly, so it was important not to excite the fluorophore too strongly or for too long. Despite this, cells stained well, showing the same actin structures seen with other phalloidin stains and had the advantage in that the product could be used more dilute (1 in 1000) compared to other stains that are used at 1 in 25 dilution. I would recommend this product if 488 or 568 fluorophores could not be used.
This image is courtesy of an anonymous Abreview.
- Fluorescence Microscopy
Supplier Data
Fluorescence Microscopy - Phalloidin-iFluor 405 Reagent (AB176752)
Excitation and emission spectra of phalloidin-iFluor 405 reagent
Reactivity data
Product details
Phalloidin-iFluor 405 Reagent (ab176752) is one of a series of phalloidin conjugates that bind to actin filaments, also known as F-actin. Phalloidin-iFluor 405 can be easily detected with a fluorescent microscope at Ex/Em = 400/421 nm.
Each 300 test product includes 1 vial containing 30 µg of iFluor 405-dye-conjugated-phalloidin in 30 µL. We recommend using this at a 1:1000 dilution, see protocol booklet for more instructions.
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.
Review other popular phalloidin dye conjugates, including Phalloidin-iFluor 488 (ab176753), Phalloidin-iFluor 647 (ab176759), Phalloidin-iFluor 594 (ab176757), Phalloidin-iFluor 555 (ab176756), and Rhodamine Phalloidin (ab235138), search the website to see all phallodin conjugates, or read the phalloidin staining protocol.
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.
Reagent Preparation
Before opening, briefly centrifuge the small vial at low speed to ensure all contents are collected at the bottom and to prevent loss of material.
Properties and storage information
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Product protocols
- Visit the General protocols
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Publications (35)
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The Journal of experimental medicine 222: PubMed39656203
2024
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Nature immunology 26:42-52 PubMed39633178
2024
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FEBS letters 599:330-339 PubMed39300480
2024
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Nature communications 15:6953 PubMed39138205
2024
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iScience 27:109825 PubMed38799552
2024
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American journal of human genetics 111:1239 PubMed38723631
2024
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American journal of human genetics 111:805 PubMed38508193
2024
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Advanced functional materials 34: PubMed38465199
2024
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American journal of human genetics 111:96-118 PubMed38181735
2024
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iScience 26:107361 PubMed37554445
2023
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Product promise
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