Overview

  • Product name
    Phalloidin-iFluor 405 Reagent
    See all F-actin kits
  • Sample type
    Adherent cells, Suspension cells
  • Assay type
    Cell-based (qualitative)
  • Product overview

    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.


    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 our other popular phalloidin dye conjugates, including Phalloidin-iFluor 488Phalloidin-iFluor 647Phalloidin-iFluor 594Phalloidin-iFluor 555, and Rhodamine Phalloidin, or search our website to see the rest of the range.

  • Notes

    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.

  • Platform
    Fluorescence microscope

Properties

Images

  • 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.

  • Excitation and emission spectra of phalloidin-iFluor 405 reagent

Protocols

References

This product has been referenced in:
  • Cervero P  et al. Lymphocyte-specific protein 1 regulates mechanosensory oscillation of podosomes and actin isoform-based actomyosin symmetry breaking. Nat Commun 9:515 (2018). Read more (PubMed: 29410425) »
  • Struchtrup A  et al. The ciliary protein RPGRIP1L governs autophagy independently of its proteasome-regulating function at the ciliary base in mouse embryonic fibroblasts. Autophagy 14:567-583 (2018). Read more (PubMed: 29372668) »
See all 5 Publications for this product

Customer reviews and Q&As

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. Actin 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.

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Verified customer

Submitted Aug 14 2015

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