Phalloidin-iFluor 488 Reagent (ab176753)

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Overview

  • Product name

    Phalloidin-iFluor 488 Reagent
    See all F-actin kits

  • Sample type

    Adherent cells, Suspension cells

  • Assay type

    Cell-based (qualitative)

  • Product overview

    Phalloidin-iFluor 488 Reagent ab176753 is one of a series of phalloidin conjugates that bind to actin filaments, also known as F-actin. The iFluor 488 dye can be easily detected with a fluorescent microscope at Ex/Em = 493/517 nm.


    Our 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 with paraffin-embedded samples that have been de-paraffinized.


    Review other popular phalloidin dye conjugates, including Phalloidin-iFluor 647, Phalloidin-iFluor 594, Phalloidin-iFluor 555, and Rhodamine Phalloidin, search the website to see all phallodin conjugates, or read the phalloidin staining protocol.

  • 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

  • Whole Mount Immunofluorescence of Mouse Inner Ear
    Whole Mount Immunofluorescence of Mouse Inner EarImage courtesy of Mr. Shahar Taiber

    Ab176753 at 1/1000 dilution in PBS whole mount Immunofluorescence of Mouse inner ear sensory epithelia. Tissue was incubated for 2 hours at room temperature.

  • Phalloidin iFluor 488 staining of macrophages
    Phalloidin iFluor 488 staining of macrophagesReview by Dr. Jose Ramos Vivas

    Macrophage J774A.1 cell line stained with Phalloidin 488 and DAPI nuclear counterstaining.

  • CytoPainter Phalloidin-iFluor 488 Reagent (ab176753)
    CytoPainter Phalloidin-iFluor 488 Reagent (ab176753)

    Actin filaments staining in HeLa cells. Actin filaments (green) were stained with CytoPainter Phalloidin-iFluor 488 reagent (ab176753); tubulin filaments were stained with a mouse anti-tubulin antibody/goat anti-mouse IgG (red). Nuclei were stained with Hoechst 33342.

  • CytoPainter Phalloidin-iFluor 488 Reagent (ab176753)
    CytoPainter Phalloidin-iFluor 488 Reagent (ab176753)

    Excitation and emission spectra of phalloidin-iFluor 488 reagent.

Protocols

References

This product has been referenced in:

  • Wang JN  et al. Acetyl-macrocalin B suppresses tumor growth in esophageal squamous cell carcinoma and exhibits synergistic anti-cancer effects with the Chk1/2 inhibitor AZD7762. Toxicol Appl Pharmacol 365:71-83 (2019). Read more (PubMed: 30633885) »
  • Hacohen-Kleiman G  et al. Atypical Auditory Brainstem Response and Protein Expression Aberrations Related to ASD and Hearing Loss in the Adnp Haploinsufficient Mouse Brain. Neurochem Res N/A:N/A (2019). Read more (PubMed: 30659505) »
See all 54 Publications for this product

Publishing research using ab176753? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-10 of 10 Abreviews or Q&A

lab Manager

 Good Good 4/5 (Ease of Use)
Abreviews
The reagent works

Abcam user community

Verified customer

Submitted Dec 27 2019

CytoPainter Phalloidin-iFluor 488 labeling of plant material

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
Detection of F-actin in live pollen of 3 species: Gagea lutea, Saintpaulia ionatha and Yucca filamentosa:

Pollen from - 20 degrees Celsius were placed in 4 degrees Celsius for a night.
Next day placed in PBS buffer for 5 min and transferred into 1xPhalloidin-iFluor 488 working solution (0,5 ul + 500 ul PBS buffer) for 20 or 90 min at RT in darkness.
Immediately visualized using Leica DM6000B with FITC filter.

Abcam user community

Verified customer

Submitted Dec 16 2019

Phalloidin-iFluor 488 Reagent

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
Bronchial epithelial cell line. 4% paraformaldehyde fixation for 15min RT. Stain with Phalloidin (1:1000) for 45 min at RT and nucleus co-stained with DAPI.

Abcam user community

Verified customer

Submitted Jul 10 2019

Good staining

 Good Good 4/5 (Ease of Use)
Abreviews
abreview image
Epithelial cells:
- Fixed 4% PFA (room temperature)
- Dilution 1:1000
- DAPI co-staining
- Fluorescence measured with confocal microscopy

Abcam user community

Verified customer

Submitted Apr 24 2019

Whole mount IF for the inner ear

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
I used abcam's phalloidin-488 for whole mount IF of mouse inner ear (sensory epithelia). It stains actin so stereocilia are stained very strong. Diluted 1:1000 in PBS and incubated for 2 hours in RT.

Mr. Shahar Taiber

Verified customer

Submitted Jun 04 2018

Phalloidin immunofluorescence of lim2099 cells

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
Seeding of 200.000 cells/well on cover slips in a 24-well-plate overnight.
5 min washing with PBS 0.1% Tween
20 min fixation with 4% PFA at room temperature.
3 x 5 min washing with PBS 0.1% Tween
10 min permeabilization with PBS 0.3% Triton-X 100
3 x 5 min washing with PBS 0.1% Tween.
Phalloidin staining for 1 hour at room temperature in the dark: 100 µl of 1x reagent diluted in PBS 0.1% Triton-X 100 1.5% BSA per coverslip.
3 x 5 min PBS 0.1% Tween.
Mounting of coverslips with ProLong Gold Antifade Mountant with DAPI on microscope slides.
Drying overnight at 4°C and imaging on the next day.

CytoPainter Phalloidin-iFluor 488 Reagent is an excellent stain yielding great results at low expenses in comparison to other more expensive stains from competing companies.

Abcam user community

Verified customer

Submitted Jun 29 2017

F-Actin / Peritoneal Macrophages

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
Excellent product for F-Actin staining. Very bright, yields minimal background if any when used correctly. Can be used for z-stacking without losing the signal in steps.

Gokhan Yilmaz

Verified customer

Submitted Mar 31 2017

CytoPainter Phalloidin-iFluor 488

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
For our experiments, the macrophage J774A.1 cell line was placed in 24-well tissue culture plates containing round glass coverslips. After 24h, cells were washed four times and fixed with cold paraformaldehyde (3.2% in PBS) for 20 min at room temperature. After being fixed, the cells were permeabilized with Triton X-100 (0.1% in PBS) for 5 min at room temperature and washed four times with PBS. CytoPainter Phalloidin-iFluor 488 (1:1000 dilution in 1%BSA-PBS), which binds polymerized F-actin, was used to identify actin filaments and fibers. Coverslips were mounted on glass slides with mounting medium containing DAPI.
Preparations were examined with a Nikon A1R confocal scanning laser microscope equipped with 405nm/450nm (DAPI) and 488nm/525nm (CytoPainter iFluor) lasers. Images were captured at random with a ×20 (A), ×40 (B) or ×60 (C, D) objectives, and processed using the NIS-Elements 3.2 software. Scale bars: A, 50µm B, 10 µm.

Dr. Jose Ramos Vivas

Verified customer

Submitted Jun 24 2016

Confocal image of MLO-Y4 cells stained with Phalloidin-iFluor 488 (& nuclear stained with DAP & sclerostin antibody labbeled with iFluor 550)

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
I stained the murine osteocyte cells (MLO-Y4 cells line) with Phalloidin-iFluor 488, DAPI staining and sclerostin antibody labbeled with iFluor 550. I used this protocol :
Fixed cells with PFA 4% for 20 min at room temperature
Wash 3 times for 5 min with PBS
Incubate for 10 min at room temperature with PBS-Triton 0.1%
Wash 3 times for 5 min with PBS-Tween 0,1%
Blocked for 30 min at room temprature with PBS-BSA 1%
Incubate with primary antibody in PBS-BSA 1% overnight at 4°C (antibody against sclerostin in this case)
Wash 3 times for 5 min with PBS-Tween 0,1%
Incubate with secondary antibody in PBS-BSA1% for 1h at room temperature in the dark (antibody anti-goat in this case)
Wash 3 times for 5 min with PBS-Tween 0,1%
Incubate with Phalloidin 488 PBS-BSA 1% 60 min at room temperature in the dark
Wash 3 times for 5 min with PBS-Tween 0,1%
Staining with DAPI for 15 min at room temprature in the dark
Wash 3 times for 5 min with PBS-Tween 0,1%
Air dry and mounting with fluorescent mounting medium

Abcam user community

Verified customer

Submitted Mar 11 2016

Good performance

 Good Excellent 5/5 (Ease of Use)
Abreviews
abreview image
Huh-7 cells: 30 min incubation including 5 min prior treatment with 0.1% Triton-X.

Dr. Teresa Otto

Verified customer

Submitted Jul 22 2015

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