Overview

  • Product name
    Phalloidin-iFluor 647 Reagent
    See all F-actin kits
  • Sample type
    Adherent cells, Suspension cells
  • Assay type
    Cell-based (qualitative)
  • Product overview

    Phalloidin-iFluor 647 Reagent (ab176759) is one of a series of phalloidin conjugates that bind to actin filaments, also known as F-actin. 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.


    Review our other popular phalloidin dye conjugates, including Phalloidin-iFluor 488Phalloidin-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

  • Fluorescence microscopy (FM) image of laminin-1 and actin (phalloidin) staining.
  • Actin filaments staining in HeLa cells. Actin filaments (red) were stained with CytoPainter Phalloidin-iFluor 647 reagent (ab176759). Nuclei were stained with DAPI (blue).

  • Excitation and emission spectra of phalloidin-iFluor 647 reagent.

Protocols

References

This product has been referenced in:
  • Burgoyne T  et al. Correlative light and immuno-electron microscopy of retinal tissue cryostat sections. PLoS One 13:e0191048 (2018). Read more (PubMed: 29315318) »
  • Curto J  et al. CK1e and p120-catenin control Ror2 function in noncanonical Wnt signaling. Mol Oncol N/A:N/A (2018). ICC/IF ; Human . Read more (PubMed: 29465811) »
See all 6 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

F-actin in Drosophila indirect flight muscle

Excellent Excellent 5/5 (Ease of Use)
Abreviews
The product stained the F-actin clearly and consistently. A 1:1000 dilution was used from 1000X stock following ABCAM instructions. The muscle tissue was imaged at 40X magnification

Dr. Daniel Babcock

Verified customer

Submitted Sep 17 2018

Actin staining of porcine MSCs

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Very easy to use. A dilution of 1:2000 was used and the F-actin was still perfectly stained.

Abcam user community

Verified customer

Submitted Apr 06 2018

STED microscopy of PFA-fixed HaCaT cells

Excellent Excellent 5/5 (Ease of Use)
Abreviews
We used Phalloidin-iFluor647 to stain Actin in HaCaT cells in STED microscopy. The reagent yielded very bright samples and the iFluor reagent worked quite nicely with STED. There was a loss in fluorescence intensity of about 60-70% compared to confocal image, but that should not be a problem in well stained samples.
The cells were fixed with 4% PFA and permeabilized with 0,2% Triton X-100. Samples were blocked with 3% BSA/PBS for 30 min. We diluted the reagent stock 1:1000 accordingly to the manufacturers instructions and added the reagent during the secondary antibody step (dilution in 3% BSA/PBS). The incubation time was 1 h at room temperature.

Mr. Membrane Biochemistry

Verified customer

Submitted Dec 04 2017

Abreviews
Rat kidneys were perfused in situ through the left heart ventricle with a fixative solution containing 3% paraformaldehyde in phosphate buffered saline (PBS), post-fixed with 1% paraformaldehyde over-night and cryoprotected with 2.3M sucrose for 12h. Cryosections 5um thick. Tissue rehydrated with PBS 30 min. Blocking with 1XPBS/ 5% donkey serum/ 0.3% Triton X-100 solution for 20 min. Phalloidin-cytoPainter 647 (ab176759) at the concentration 1:1000 (stock 1000x according with Abcam instruction) in a solution of 1XPBS/ 1% BSA/ 0.3% Triton-X-100 for 1h at room temperature protected from light. Wash 3 times with PBS.

Mr. Udo Schnitzbauer

Verified customer

Submitted Dec 03 2015

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