Fitc labeled phalloidin

Manufactured by Merck Group

Sourced in United States, Germany

FITC-labeled phalloidin is a fluorescent stain used for the detection and visualization of actin filaments in cells. It binds specifically to F-actin and can be used in various microscopy techniques to study the organization and dynamics of the actin cytoskeleton.

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Lab products found in correlation

Triton x 100, by Merck Group (4 mentions) Normal goat serum (ngs), by ZSGB-BIO (3 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (3 mentions) Normal goat serum (ngs), by Merck Group (3 mentions) Paraformaldehyde (pfa), by Merck Group (3 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (3 mentions) Lsm 780 confocal microscope, by Zeiss (3 mentions) Fluorescence microscope, by Leica (2 mentions) Dapi 4 6 diamidino 2 phenylindole, by Merck Group (2 mentions) Axiovert 700 microscope, by Zeiss (2 mentions) Vectashield mounting medium, by Vector Laboratories (2 mentions) Perm wash buffer, by BD (2 mentions) Lsm 780, by Zeiss (2 mentions) Paraformaldehyde (pfa), by Electron Microscopy Sciences (2 mentions) Lsm 510 meta confocal microscope, by Zeiss (2 mentions) Fitc phalloidin, by Merck Group (1 mentions) Cytofix cytoperm buffer, by BD (1 mentions) Facscalibur, by BD (1 mentions) Cell chamber slides, by Corning (1 mentions) Fv1000 confocal microscope, by Olympus (1 mentions)

Spelling variants of this product

FITC-phalloidin (327 citations) Fluorescein isothiocyanate (FITC)-labeled phalloidin (3 citations) Fluorescein isothiocyanate (FITC)-labeled phalloidin (green fluorescence) (2 citations)

47 protocols using fitc labeled phalloidin

Quantifying Cellular F-actin Content

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F-actin staining using FITC-phalloidin (Sigma-Aldrich, St. Louis, MO) was carried out using 1 × 10⁶ cells. The cells were pelleted, fixed, and permeabilized with CytoPerm/Cytofix buffer (BD Biosciences, San Jose, CA) for 20 min at room temperature and washed with cold Perm/Wash buffer (BD Biosciences, San Jose, CA) twice, followed by staining with 5 μl of 0.3 mM FITC-labeled phalloidin (Sigma-Aldrich, St. Louis, MO) for 30 min on ice in the dark. The cells were washed twice with cold Perm/Wash buffer, resuspended in 1% paraformaldehyde, and analyzed on a FACSCalibur (BD Biosciences, San Jose, CA).

Yi F., Guo J., Dabbagh D., Spear M., He S., Kehn-Hall K., Fontenot J., Yin Y., Bibian M., Park C.M., Zheng K., Park H.J., Soloveva V., Gharaibeh D., Retterer C., Zamani R., Pitt M.L., Naughton J., Jiang Y., Shang H., Hakami R.M., Ling B., Young J.A., Bavari S., Xu X., Feng Y, & Wu Y. (2017). Discovery of Novel Small-Molecule Inhibitors of LIM Domain Kinase for Inhibiting HIV-1. Journal of Virology, 91(13), e02418-16.

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F-actin Visualization in Endothelial Cells

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HUVECs were seeded onto glass coverslips placed in 24-well plates till growth achieved confluence. Anti-ATP5B antibodies (50 μg/ml) were added for 1 h, followed by FlgE or FlgEM (20 μg/ml) challenge for 2 h. Cells were washed with pre-warmed PBS and fixed with 4% PFA for 20 min at 37°C. The cells were then permeabilized with 0.5% Triton X-100 for 15 min and blocked for 1 h with 3% BSA in PBS at room temperature. For F-actin staining, 5 μg/ml of FITC-labeled phalloidin (Sigma) was added for 40 min. Nuclei were counterstained with DAPI. After washing, the coverslips were observed with a fluorescence microscope (Leica), and images were captured.

Li Y., Shen Y., Zheng Y., Ji S., Wang M., Wang B., Han Q., Tian Y, & Wang Y. (2021). Flagellar Hook Protein FlgE Induces Microvascular Hyperpermeability via Ectopic ATP Synthase β on Endothelial Surface. Frontiers in Cellular and Infection Microbiology, 11, 724912.

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Immunofluorescence Staining of MDA-MB-231 Cells

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MDA-MB-231 cells (2×10⁴) were seeded onto cell chamber slides (Corning, Tewksbury, MA, USA) in 24-well plates 24 h before assaying. The cells on the slides were fixed with 4% formaldehyde and then stained by FITC-labeled Phalloidin (Sigma-Aldrich, St. Louis, MO, USA, 1: 500 dilution), or stained with anti-LMO2 and anti-ARP3/anti-profilin1 antibodies (1: 200 dilution) at 4°C overnight, followed by incubating with the appropriate fluorescent secondary antibodies at room temperature for 1 h. Images were captured using an FV1000 confocal microscope (Olympus, Center Valley, PA, USA).

Liu Y., Wu C., Zhu T, & Sun W. (2017). LMO2 Enhances Lamellipodia/Filopodia Formation in Basal-Type Breast Cancer Cells by Mediating ARP3-Profilin1 Interaction. Medical Science Monitor : International Medical Journal of Experimental and Clinical Research, 23, 695-703.

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Immunostaining of Fibronectin Matrix

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Fixed samples were washed three times in PBS, and permeabilized using 0.1% Triton-X in PBS for 20 minutes at room temperature. Fibronectin (FN) matrix proteins were fluorescently labeled using standard indirect immunostaining protocols. The samples were blocked with 3% bovine serum albumen (BSA) for 30 minutes at 37 °C, and incubated overnight at 4 °C in a humidified chamber with a 1:200 dilution of anti-FN primary antibodies in 0.1% BSA solution. The samples were then washed three times in PBS, blocked with 10% goat serum for 30 minutes at room temperature, and incubated with a secondary Alexafluor 488 antibody (Invitrogen) for 1 hour at room temperature. Direct labeling for actin fibers was conducted by incubating samples with 0.1 μM FITC-labeled phalloidin (Sigma) in 0.1% BSA for 30 minutes at room temperature. Nuclei were labeled with Hoechst 33258 (10 μg/mL in 0.1% BSA) for 15 minutes at room temperature. In all cases, the samples were washed in PBS thoroughly, and mounted on a glass coverslip using Fluoromount G (Southern Biotech).
The samples were imaged using either epifluorescent (TE300, Nikon) or confocal (Leica SP5) microscopy. Image reconstruction and analysis were conducted in ImageJ (NIH). Nuclear dimensions were calculated by fitting ellipses to nuclear images using native ImageJ functions, and extracting dimensions of long and short axes.

Moraes C., Kim B.C., Zhu X., Mills K.L., Dixon A.R., T.h.o.u.l.e.s.s, & Takayama S. (2014). Defined topologically-complex protein matrices to manipulate cell shape via three-dimensional fiber-like patterns. Lab on a chip, 14(13), 2191-2201.

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PBMC F-actin Polymerization Assay

Cited 1 time

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Isolated PBMCs were cultured for 12 hours in R-10 before the F-actin polymerization test. The cells were first stained with PE-conjugated anti-CD45R/B220 mAb to discriminate between B- and T-cell populations before being subjected to the F-actin polymerization test. Intracellular F-actin polymerization was assessed as previously described [31 (link)]. Briefly, the cells were harvested and resuspended (4 x 10⁶/ml) in HEPES-buffered RPMI-1640 at 37°C with or without CXCL12 (500 ng/ml). At the indicated times, the cell suspensions (100 μl) were added to 400 μl of assay buffer containing 4 x 10⁷ FITC-labeled phalloidin, 0.5 mg/ml L-α-lysophosphatidylcholine (both from Sigma-Aldrich) and 4.5% formaldehyde in PBS. The fixed cells were analyzed by flow cytometry, and the mean fluorescence intensity (MFI) was determined for each sample. The percent change in MFI was calculated for each sample at each time point according to the following formula: (1-(MFI before the addition of CXCL12/MFI after the addition of CXCL12)) × 100.

Badr G., Sayed A., Abdel-Maksoud M.A., Mohamed A.O., El-Amir A., Abdel-Ghaffar F.A., Al-Quraishy S, & Mahmoud M.H. (2015). Infection of Female BWF1 Lupus Mice with Malaria Parasite Attenuates B Cell Autoreactivity by Modulating the CXCL12/CXCR4 Axis and Its Downstream Signals PI3K/AKT, NFκB and ERK. PLoS ONE, 10(4), e0125340.

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Insulin-Loaded Mucoadhesive Nanoparticles

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2-(N,N'-dimethylamino) ethyl methacrylate (DMAEMA, 98%) was purchased from Alfa Aesar. β-Propiolactone (98%) and 2-cyanoprop-2-yl-dithiobenzoate were from J&K Scientific Ltd. Porcine insulin, FITC-labeled insulin and trypsin were purchased from Dalian Meilun Biotechnology Co., Ltd. 2,2'-Dicyano-2,2'-azopropane (AIBN) was from Aladdin Industrial Corporation. MTT, FITC-labeled phalloidin, porcine stomach mucin and STZ were obtained from Sigma Aldrich. LysoTracker deep and Pierce^TM Coomassie Plus (Bradford) Assay Kit were purchased from Thermo Fisher Scientific. Alamar blue assay, 4% paraformaldehyde, normal goat serum and bafilomycin A1 were from Beijing Solarbio Science & Technology Co., Ltd. The Eudragit L 100-55 was from Shanghai Chineway Pharmaceutical Technology Co., Ltd. Triton® X-100 was from MP Biomedicals, LLC. Anti-claudin-4 antibody and Alexa-Fluor-conjugated secondary antibody were obtained from Abcam. Anti-integrin αV and β1 antibodies were purchased from Affinity Biosciences LTD. All plates used in cellular experiments were Corning Incorporated.

Li Y., Ji W., Peng H., Zhao R., Zhang T., Lu Z., Yang J., Liu R, & Zhang X. (2021). Charge-switchable zwitterionic polycarboxybetaine particle as an intestinal permeation enhancer for efficient oral insulin delivery. Theranostics, 11(9), 4452-4466.

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Immunostaining of EGFR and E-cadherin

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Cells used for immunostaining were fixed in ice-cold methanol for 10 min, permeabilized in 0.1 % Triton X-100 and blocked in PBS containing 1 % BSA for 1 h at room temperature. The cells were incubated with antibody against EGFR or E-cadherin at 4 °C overnight followed by incubation with rhodamine-conjugated or FITC-conjugated secondary antibody for 1 h at room temperature within a moist chamber. F-actin was stained with FITC-labeled phalloidin (5 μg/mL) (Sigma) for 45 min at 37 °C. After wash with PBS, the samples were mounted with DAPI Fluoromount G (Southern Biotech, Birmingham, AL). Images were acquired using an Olympus BX51 microscope coupled with an Olympus DP70 digital camera.

Deng W., Gu L., Li X., Zheng J., Zhang Y., Duan B., Cui J., Dong J, & Du J. (2016). CD24 associates with EGFR and supports EGF/EGFR signaling via RhoA in gastric cancer cells. Journal of Translational Medicine, 14, 32.

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Antibody Characterization Protocol

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Rabbit anti–TIMP-1 polyclonal antibody (pAb) was purchased from Abcam (Cambridge, USA). Mouse anti–CD82 monoclonal antibody (mAb), anti–GFP mAb, anti–α-tubulin mAb, and anti–β-Actin mAb were purchased from Santa Cruz Biotechnology. Stabilized Streptavidin-HRP Conjugate was obtained from Thermo Scientific. We purchased FITC-labeled phalloidin from Sigma (St. Louis, USA). We obtained horseradish peroxidase-conjugated secondary antibodies from Amersham Pharmacia Biotech (Piscataway, USA). Fluorescent-labeled secondary antibodies were purchased from Invitrogen.

Zhang J., Wu T., Zhan S., Qiao N., Zhang X., Zhu Y., Yang N., Sun Y., Zhang X.A., Bleich D, & Han X. (2016). TIMP-1 and CD82, a promising combined evaluation marker for PDAC. Oncotarget, 8(4), 6496-6512.

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Autophagy Regulation via CXCR4 and IL-6 Signaling

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Autophagy inducer PP242 was purchased from Selleck chemicals (Houston, TX). Anti-CXCR4-APC (eBioscience, San Diego, CA, anti-VLA-4-APC (BD Bioscience, San Jose, CA) and anti-VCAM-1-APC (Biolegend, San Diego, CA) were used for FACs analysis. The following antibodies were used for Western blot: anti-Gli1 (Cell Signaling, Danvers, MA), anti-Gli2 (Santa Cruz, Dallas, TX), anti-Ptch1 (Santa Cruz, Dallas, TX), anti-FAK (AbCam, Cambridge, MA), anti-phospho-FAK (Cell Signaling, Danvers, MA), anti-paxillin (BD Biosciences, San Jose, CA), anti-phospho-paxillin (Cell Signaling, Danvers, MA), anti-SDF-1 (Santa Cruz, Dallas, TX), anti-IL-6 (R&D Systems, Minneapolis, MN), anti-LC3 (Novus Biologicals, Littleton, CO), and anti-GAPDH (Cell Signaling, Danvers, MA). Recombinant human SDF-1 (rhSDF-1) was purchased from Pepro Tech (Rocky Hill, NJ), and recombinant human IL-6 (rhIL-6) was purchased from R&D System (Minneapolis, MN). CXCR4 antagonist AMD3100, 2,7-dichlorodihydrofluorescein-diacetate (DCH-FDA, 50MG), ROS inhibitor N-acetyl-L-cysteine (NAC), FITC-labeled phalloidin, l-alpha-lysophosphatidylcholine, and autophagy inhibitor 3-MA were all purchased from Sigma-Aldrich (St Louis, MO).

Zhang H., Chen Z., Neelapu S.S., Romaguera J, & McCarty N. (2016). Hedgehog inhibitors selectively target cell migration and adhesion of mantle cell lymphoma in bone marrow microenvironment. Oncotarget, 7(12), 14350-14365.

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Osteogenic Differentiation of hMSCs

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After 21 and 35 days hMSCs cultured on the 3D scaffold or the 2D substrates either in BM or OS (n = 4) were fixed with 3% paraformaldehyde (PFA) solution containing 2% sucrose for 10 min at room temperature (RT) and rinsed with PBS twice. The cells were then stained with 1:200 diluted FITC-labeled phalloidin (Sigma) for 20 min at 37 °C, followed by 1:800 diluted Hoechst 33258 (Sigma) staining for 10 min at RT. Afterwards, the fluorescent signals were observed using a confocal laser-scanning microscope (Zeiss LSM 780, Oberkochen, Germany). The cell proliferation and cell viability were measured using a MTT (3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide) assay at 7, 21 and 35 days of cell culture in BM. The cell culture medium was completely replaced with new medium containing MTT (0.1 mg/ml medium) and the test samples were incubate at 37 °C for 3 h. Unincorporated dye was removed, dimethyl-sulfoxide (DMSO) was added and absorbance was quantified by plate reader (Victor 2, PerkinElmer Life Science/Wallac Oy, Turku, Finland) at 550 nm. The result was background-corrected at an absorbance of 650 nm. The results were expressed as the average absorbance values of four replicates.

Persson M., Lehenkari P.P., Berglin L., Turunen S., Finnilä M.A., Risteli J., Skrifvars M, & Tuukkanen J. (2018). Osteogenic Differentiation of Human Mesenchymal Stem cells in a 3D Woven Scaffold. Scientific Reports, 8, 10457.

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