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Alexa fluor 568 coupled phalloidin

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Alexa Fluor 568-coupled phalloidin is a fluorescently-labeled version of the natural product phalloidin. Phalloidin binds to and stabilizes filamentous actin (F-actin) in cells, which allows for the visualization of the actin cytoskeleton.

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Lab products found in correlation

Anti fade with dapi, by Thermo Fisher Scientific (2 mentions) Sp5 confocal microscope, by Leica (2 mentions) Fitc coupled gelatine, by Thermo Fisher Scientific (2 mentions) Bz 9000 microscope, by Keyence (1 mentions) Triton x 100, by Merck Group (1 mentions) Trypsin edta, by Merck Group (1 mentions) Mouse anti gapdh, by Merck Group (1 mentions) Irdye 680rd goat anti mouse, by LI COR (1 mentions) Recombinant human tgf β1 protein, by R&D Systems (1 mentions) Rabbit anti zo 1 antibody, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Rabbit anti fibronectin antibody, by Merck Group (1 mentions) Mouse anti vimentin antibody, by Merck Group (1 mentions) Rabbit anti p smad3, by Cell Signaling Technology (1 mentions) Mouse anti smad2 3, by BD (1 mentions) Mouse anti paxillin antibody, by BD (1 mentions)

Spelling variants of this product

Alexa Fluor 568 (1190 citations) Alexa Fluor 568 phalloidin (436 citations) Alexa 568 (337 citations) Alexa Fluor phalloidin (43 citations) Phalloidin 568 (31 citations) Alexa Fluors (21 citations) Phalloidin-Alexa-568 (8 citations) Phalloidins (2 citations)

4 protocols using alexa fluor 568 coupled phalloidin

1

Quantification of Matrix Degradation

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Cover slips were coated with 0.2 mg/ml FITC-coupled gelatine (G13187, Molecular Probes) and subsequently blocked using complete media. The siRNA-treated cells were seeded on the cover slips and incubated at 37°C for 4 hours and fixed with 4% PFA. The cells were labelled with Alexa Fluor 568-coupled phalloidin (A12380, Molecular Probes) and mounted in anti-fade with DAPI (P36931, Molecular Probes). Cells were imaged on a Leica SP5 confocal microscope. The images were processed using ImageJ (Wayne Rasband, NIH). Matrix degradation was quantified by measuring the area where the FITC labelled-gelatine was below the threshold and counting the number of cells using ImageJ, as described before31 .
Curtis J., Luo Y., Zenner H.L., Cuchet-Lourenço D., Wu C., Lo K., Maes M., Alisaac A., Stebbings E., Liu J.Z., Kopanitsa L., Ignatyeva O., Balabanova Y., Nikolayevskyy V., Baessmann I., Thye T., Meyer C.G., Nürnberg P., Horstmann R.D., Drobniewski F., Plagnol V., Barrett J.C, & Nejentsev S. (2015). Susceptibility to tuberculosis is associated with variants in the ASAP1 gene encoding a regulator of dendritic cell migration. Nature genetics, 47(5), 523-527.
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2

Quantification of Matrix Degradation

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Cover slips were coated with 0.2 mg/ml FITC-coupled gelatine (G13187, Molecular Probes) and subsequently blocked using complete media. The siRNA-treated cells were seeded on the cover slips and incubated at 37°C for 4 hours and fixed with 4% PFA. The cells were labelled with Alexa Fluor 568-coupled phalloidin (A12380, Molecular Probes) and mounted in anti-fade with DAPI (P36931, Molecular Probes). Cells were imaged on a Leica SP5 confocal microscope. The images were processed using ImageJ (Wayne Rasband, NIH). Matrix degradation was quantified by measuring the area where the FITC labelled-gelatine was below the threshold and counting the number of cells using ImageJ, as described before31 .
Curtis J., Luo Y., Zenner H.L., Cuchet-Lourenço D., Wu C., Lo K., Maes M., Alisaac A., Stebbings E., Liu J.Z., Kopanitsa L., Ignatyeva O., Balabanova Y., Nikolayevskyy V., Baessmann I., Thye T., Meyer C.G., Nürnberg P., Horstmann R.D., Drobniewski F., Plagnol V., Barrett J.C, & Nejentsev S. (2015). Susceptibility to tuberculosis is associated with variants in the ASAP1 gene encoding a regulator of dendritic cell migration. Nature genetics, 47(5), 523-527.
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3

Imaging Actin Cytoskeleton in AML Cells

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AML cells grown on poly-L-lysin coated chamber slides (Ibidi) were fixed with 4% paraformaldehyd/PBS for 10 min 37 °C. After washing the cells 3-times with PBS, they were incubated with Alexa-fluor 568-coupled phalloidin (Thermo Fisher Scientifics, Waltham, MA, USA), diluted 1:1000 in PBS, for 30 min at RT. Thereafter, the cells were washed again 3-times with PBS and stained with 4′,6-Diamidin-2-phenylindo (1:2000 in PBS) for 5 min at RT. After final washing, fluorescence was analyzed by the Keyence BZ 9000 microscope.
Velthaus A., Cornils K., Hennigs J.K., Grüb S., Stamm H., Wicklein D., Bokemeyer C., Heuser M., Windhorst S., Fiedler W, & Wellbrock J. (2019). The Actin Binding Protein Plastin-3 Is Involved in the Pathogenesis of Acute Myeloid Leukemia. Cancers, 11(11), 1663.
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4

Immunofluorescence Analysis of Cell Adhesion

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The mouse anti-paxillin antibody was purchased from BD Biosciences (610052) and the corresponding secondary Alexa488 anti-mouse antibody from ThermoFisher Scientific (A-11029). The rabbit anti-fibronectin antibody was obtained from Sigma (F3648) and the corresponding secondary Alexa647 anti-rabbit antibody from ThermoFisher Scientific (A-21245). Alexa Fluor 568-coupled phalloidin was from ThermoFisher Scientific (A12380). Rat anti-E-cadherin antibody was obtained from ThermoFisher Scientific (13-1900), rabbit anti-ZO1 antibody from ThermoFisher Scientific (61-7300), mouse anti-vimentin antibody from Sigma (V2258), mouse anti-SMAD2/3 from BD Biosciences (610842), rabbit anti-p-SMAD3 from Cell Signaling (p-Ser423/425; 9520) and mouse anti-GAPDH from Sigma (G8795). For quantitative immunoblot analysis, secondary antibodies from LI-COR Biosciences, IRDye 680RD Goat anti-Mouse (926-68070) and IRDye 800CW Goat anti-Rabbit (926-32211) were used. DAPI was acquired from Sigma (D9542), recombinant human TGFβ1 protein from R & D Systems (240B-0-10), 16% paraformaldehyd (PFA) from Electron Microscopy Services (15710-S), fatty-acid free BSA from Calbiochem (126575), Trypsin/EDTA from Sigma (T4174), PBS from Gibco (14200-067), and Triton X-100 from Sigma (X-100).
Lotz-Jenne C., Lüthi U., Ackerknecht S., Lehembre F., Fink T., Stritt M., Wirth M., Pavan S., Bill R., Regenass U., Christofori G, & Meyer-Schaller N. (2016). A high-content EMT screen identifies multiple receptor tyrosine kinase inhibitors with activity on TGFβ receptor. Oncotarget, 7(18), 25983-26002.
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