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Af 488 conjugated phalloidin

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AF 488-conjugated phalloidin is a fluorescent probe that selectively binds to filamentous actin (F-actin) in cells. It can be used to visualize the actin cytoskeleton in fixed and permeabilized cells.

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Lab products found in correlation

Du897, by Oxford Instruments (2 mentions) Anti rat af488, by Thermo Fisher Scientific (1 mentions) Anti rabbit af568, by Thermo Fisher Scientific (1 mentions) Streptavidin af647, by Thermo Fisher Scientific (1 mentions) Af568 conjugated phalloidin, by Thermo Fisher Scientific (1 mentions) Anti rabbit af488, by Thermo Fisher Scientific (1 mentions) Anti pkb t308 clone c25e6, by Cell Signaling Technology (1 mentions)

4 protocols using af 488 conjugated phalloidin

1

Imaging Flow Cytometry of AFM13 on NK Cells

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For immunofluorescence detection of AFM13 on the surface of NK cells, CB-NK cells were loaded with AFM13 for 1h, washed, and cultured for another 24 hours to 72 hours at 37°C in the presence of IL-2, harvested, and stained with rat anti-AFM13 (clone 7, Affimed), followed by staining with goat anti-rat IgG (Alexa Fluor 647, Jackson ImmunoReseaech). Images were obtained using imaging flow cytometry (Amnis Image Stream Mark II) and analyzed by the Ideas software (Luminex). The retention of AFM13 on the surface of NK cells was determined by the co-localization of CD16 and AFM13 signals. Imaging flow cytometry (Amnis Image Stream Mark II) was also used to assess immunological synapse formation between AFM13-loaded CB-NK cells and Karpas 299 by staining the cells with a biotinyalated anti-pericentrin primary antibody (Novusbio) followed by streptavidin-conjugated APC-Cy7 (Biolegend) and AF 488-conjugated phalloidin (Thermofisher) to measure F-actin. Images were analyzed with the IDEAS software (Miltenyi Biotech).
Kerbauy L.N., Marin-Agudelo N., Kaplan M., Banerjee P.P., Berrien-Elliott M.M., Becker-Hapak M., Basar R., Foster M., Melo L.G., Neal C., McClain E., Daher M., Cortes A.K., Desai S., Lim F.W., Mendt M.C., Schappe T., Li L., Shaim H., Shanley M., Ensley E.L., Uprety N., Wong P., Liu E., Ang S.O., Cai R., Nandivada V., Mohanty V., Miao Q., Shen Y., Baran N., Fowlkes N., Chen K., Muniz-Feliciano L., Champlin R.E., Nieto Y.L., Koch J., Treder M., Fischer W., Okamoto O.K., Shpall E.J., Fehniger T.A, & Rezvani K. (2021). Combining AFM13, a bispecific CD30/CD16 antibody, with cytokine-activated cord blood-derived NK cells facilitates CAR-like responses against CD30+ malignancies. Clinical cancer research : an official journal of the American Association for Cancer Research, 27(13), 3744-3756.
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2

Comprehensive Immunophenotyping Approach

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Anti-HSP90 (clone 3H3C27), anti-SHIP1 (clone PICI-A5), FITC-conjugated rat anti-mouse GR1 (clone RB6-8C5), PE-conjugated rat anti-mouse/human CD11b (clone M1/70), PE/Cy7-conjugated rat anti mouse/human CD45 (clone 30-F11), PerCP/Cy5.5-conjugated rat anti F4/80 (clone BM8), APC-conjugated rat anti-B220 (clone RA3-6B2), PE/Cy7-conjugated rat anti CD3 (clone 17A2) and pacific blue-conjugated rat anti-LY6G (clone 1A8) were from BioLegend (London, UK); anti-PTEN (clone D4.3), anti-PKB (clone 11E7), anti-PKB T308 (clone C25E6) and anti-PKB S473 (clone D9E) were from Cell Signaling Technology (London, UK). Rabbit IgG (I8140) was obtained from Sigma. Anti-SHIP2 (AF5389) and PE-conjugated rat anti-CD64 (clone FAB20741P) were from R&D Systems (Abingdon, UK) and biotinylated anti-PI(3,4)P2 (z-B034) was from Echelon Biosciences (Salt Lake City, UT, USA); streptavidin-AF647, AF488-conjugated phalloidin, AF568-conjugated phalloidin, and secondary antibodies anti-rat AF488, anti-rabbit AF568 and anti-rabbit AF488 were obtained from Thermo Fisher Scientific (Loughborough, UK).
Michael M., McCormick B., Anderson K.E., Karmakar U., Vermeren M., Schurmans S., Amour A, & Vermeren S. (2021). The 5-Phosphatase SHIP2 Promotes Neutrophil Chemotaxis and Recruitment. Frontiers in Immunology, 12, 671756.
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3

Visualizing Immune Synapse Formation

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Immunological synapse formation was analyzed in response to lipid bilayer stimulation as previously described (Guy et al., 2013 (link)). Briefly, lipid bilayers containing ICAM-1 and Alexa Fluor (AF)647-labeled anti-TCR antibodies were prepared. Resting OT-I CD8+ T cells were stimulated for 30 min before fixation with 4% paraformaldehyde, permeabilization with 0.1% T-100, and staining with antibodies overnight at 4°C. AF568-conjugated secondary antibodies and AF488-conjugated phalloidin (Fisher) were applied for 1 hr before analysis using TIRF microscopy. Images were acquired using an inverted TiE Nikon microscope equipped with a 100× 1.45 numerical aperture (NA) oil objective, motorized TIRF illumination, Andor DU-897 high-speed electron-multiplying charge-coupled device (EMCCD) camera, and Agilent laser launch. Analysis of fluorescent intensities for each channel were determined for individual pixels using Nikon Elements software.
Menk A.V., Scharping N.E., Moreci R.S., Zeng X., Guy C., Salvatore S., Bae H., Xie J., Young H.A., Wendell S.G, & Delgoffe G.M. (2018). Early TCR Signaling Induces Rapid Aerobic Glycolysis Enabling Distinct Acute T Cell Effector Functions. Cell reports, 22(6), 1509-1521.
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4

Visualizing Immune Synapse Formation

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Immunological synapse formation was analyzed in response to lipid bilayer stimulation as previously described (Guy et al., 2013 (link)). Briefly, lipid bilayers containing ICAM-1 and Alexa Fluor (AF)647-labeled anti-TCR antibodies were prepared. Resting OT-I CD8+ T cells were stimulated for 30 min before fixation with 4% paraformaldehyde, permeabilization with 0.1% T-100, and staining with antibodies overnight at 4°C. AF568-conjugated secondary antibodies and AF488-conjugated phalloidin (Fisher) were applied for 1 hr before analysis using TIRF microscopy. Images were acquired using an inverted TiE Nikon microscope equipped with a 100× 1.45 numerical aperture (NA) oil objective, motorized TIRF illumination, Andor DU-897 high-speed electron-multiplying charge-coupled device (EMCCD) camera, and Agilent laser launch. Analysis of fluorescent intensities for each channel were determined for individual pixels using Nikon Elements software.
Menk A.V., Scharping N.E., Moreci R.S., Zeng X., Guy C., Salvatore S., Bae H., Xie J., Young H.A., Wendell S.G, & Delgoffe G.M. (2018). Early TCR Signaling Induces Rapid Aerobic Glycolysis Enabling Distinct Acute T Cell Effector Functions. Cell reports, 22(6), 1509-1521.
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