Fluorescent phalloidin conjugate

Manufactured by Merck Group

Sourced in United States

Fluorescent phalloidin conjugate is a laboratory reagent used for the detection and visualization of F-actin, a key structural component of the cytoskeleton in eukaryotic cells. It is a fluorescently labeled derivative of the toxin phalloidin, which binds specifically to F-actin. The fluorescent label allows for the localization and quantification of F-actin within cells or tissue samples using techniques such as fluorescence microscopy.

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Lab products found in correlation

β catenin antibody, by Cell Signaling Technology (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) Crystal violet, by Thermo Fisher Scientific (1 mentions) Transwell chamber, by Corning (1 mentions) Tcs sp8 confocal microsystems, by Leica (1 mentions) Prolong gold antifade reagent with dapi, by Cell Signaling Technology (1 mentions)

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Fluorescent phalloidin (8 citations)

3 protocols using fluorescent phalloidin conjugate

Immunofluorescence Analysis of Cell Structure

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Cells were fixed in 4% formaldehyde, stained with fluorescent phalloidin conjugate (Sigma‐Aldrich) and counterstained with 4’,6’‐diamidino‐2‐phenylindole (DAPI; Invitrogen, Carlsbad, CA, USA). Images were acquired with an Olympus BX51.
Cells plated on coverslips were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X‐100 at 4 °C for 10 min. After blocking, the cells were stained with β‐catenin antibody (Cell Signaling Technology, Danvers, MA, USA) and counterstained with DAPI (Invitrogen, OR). Images were captured with a confocal microscope Olympus FV1000 or Olympus BX51.

Zeng T., Tang Z., Liang L., Suo D., Li L., Li J., Yuan Y., Guan X, & Li Y. (2020). PDSS2‐Del2, a new variant of PDSS2, promotes tumor cell metastasis and angiogenesis in hepatocellular carcinoma via activating NF‐κB. Molecular Oncology, 14(12), 3184-3197.

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Assessing Cell Migration and Invasion

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A wound-healing assay was performed using 2.5×10⁵ cells seeded into a 24-well plate. The cells were cultured with serum-reduced medium (1% FBS). After 24 hours, the wells were scraped with 200-μL tips. We recorded the wound widths immediately using a microscope and again after 48 hours for C4-2B and LNCaP, and 24 hours for PC3. Transwell assays were performed by seeding 5×10⁴ cells into transwell chambers (Costar, Cambridge, MA, USA) with 5-μm-pore polycarbonate filters. The cells were cultured using serum-reduced medium (1% FBS) for 48 hours, and the cells attached to the chamber membrane were fixed in formalin and stained with 0.05% crystal violet (Invitrogen). Cells on the top of cylindrical chambers were removed using cotton swabs. Invasive cells were determined by detecting the O.D. 540 in a plate reader. For cytoskeleton staining, 1×10⁴ cells were seeded in 96-well plate, fixed with 3.7% formaldehyde for 5 minutes, permeabilized with 0.1% TRITON-X-100 and then stained with a 50 μg/mL fluorescent phalloidin conjugate (Sigma, St. Louis, MO, USA) for 40 minutes at room temperature. Lamellipodia positive cells were imaged microscopically.

Yang J., Lu C., Wei J., Guo Y., Liu W., Luo L., Fisch G, & Li X. (2016). Inhibition of KPNA4 Attenuates Prostate Cancer Metastasis. Oncogene, 36(20), 2868-2878.

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Visualizing Actin Cytoskeleton Dynamics

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H-HSC were treated with incremental concentrations of S3I-201 for 24 h. Cells were then fixed with 4% paraformaldehyde for 5 min at room temperature, and then permeabilized with 0.1% triton X-100 for another 5 min. After washing, cells were stained with 50 μg/ml fluorescent phalloidin conjugate (Sigma, St Louis, MO, USA) at room temperature for 40 min, then washed 4-5 times with PBS, and covered with ProLong Gold Antifade Reagent with DAPI (Cell Signaling Technology, Danvers, MA, USA). Actin cytoskeleton was observed using Leica TCS SP8 confocal microsystems (Wetzlar, Germany).

Wang Z., Li J., Xiao W., Long J, & Zhang H. (2018). The STAT3 inhibitor S3I-201 suppresses fibrogenesis and angiogenesis in liver fibrosis. Laboratory investigation; a journal of technical methods and pathology, 98(12).

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