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Alexfluor550

Manufactured by Thermo Fisher Scientific
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AlexaFluor550 is a fluorescent dye produced by Thermo Fisher Scientific. It is designed for use in biological imaging and labeling applications. The dye exhibits bright fluorescence with excitation and emission maxima at 553 nm and 568 nm, respectively.

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Lab products found in correlation

Confocal microscope, by Nikon (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions) Lamp1, by Thermo Fisher Scientific (2 mentions) Myd88, by Thermo Fisher Scientific (2 mentions) Confocal dishes, by MatTek (2 mentions) Alexa fluor 488, by Thermo Fisher Scientific (2 mentions) Antibodies to mouse tlr7, by Thermo Fisher Scientific (2 mentions)

2 protocols using alexfluor550

1

Intracellular Localization of TLR7 Signaling Components in BMDCs

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BMDCs were seeded in confocal dishes (Mattek) at a final concentration of 1×105 cells/mL in RPMI-complete media. The following day, cells were treated with 10 μg/mL GRD for 5 minutes and 15 minutes at 37C. Cells were carefully washed with PBS and fixed with 2% PFA for 10 minutes at room temperature in the dark. Fixed cells were permeabilized with SAP buffer (2% saponin in PBS) for 1 hour at 37C. Following permeabilization, BMDCs were intracellularly stained with antibodies to mouse TLR7, MyD88, EEA1, or LAMP1 (Thermo Scientific), followed by staining with secondary antibodies fluorescently labeled with AlexaFluor488 or AlexFluor550 (Invitrogen). Cells were washed 3 times with PBS. Finally, DNA was stained with 1:1000 dilution of DAPI (Invitrogen) for 10 minutes at RT. Cells were visualized using a Nikon confocal microscope and analysis performed using ZEN software. Data analysis and processing of images was conducted using ImageJ software (NIH).
Ramirez-Ortiz Z.G., Prasad A., Griffith J.W., Pendergraft WF I.I.I., Cowley G.S., Root D.E., Tai M., Luster A.D., Khoury J.E., Hacohen N, & Means T.K. (2015). TREML4 amplifies TLR7-mediated signaling during antiviral responses and autoimmunity. Nature immunology, 16(5), 495-504.
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2

Intracellular Localization of TLR7 Signaling Components in BMDCs

Check if the same lab product or an alternative is used in the 5 most similar protocols
BMDCs were seeded in confocal dishes (Mattek) at a final concentration of 1×105 cells/mL in RPMI-complete media. The following day, cells were treated with 10 μg/mL GRD for 5 minutes and 15 minutes at 37C. Cells were carefully washed with PBS and fixed with 2% PFA for 10 minutes at room temperature in the dark. Fixed cells were permeabilized with SAP buffer (2% saponin in PBS) for 1 hour at 37C. Following permeabilization, BMDCs were intracellularly stained with antibodies to mouse TLR7, MyD88, EEA1, or LAMP1 (Thermo Scientific), followed by staining with secondary antibodies fluorescently labeled with AlexaFluor488 or AlexFluor550 (Invitrogen). Cells were washed 3 times with PBS. Finally, DNA was stained with 1:1000 dilution of DAPI (Invitrogen) for 10 minutes at RT. Cells were visualized using a Nikon confocal microscope and analysis performed using ZEN software. Data analysis and processing of images was conducted using ImageJ software (NIH).
Ramirez-Ortiz Z.G., Prasad A., Griffith J.W., Pendergraft WF I.I.I., Cowley G.S., Root D.E., Tai M., Luster A.D., Khoury J.E., Hacohen N, & Means T.K. (2015). TREML4 amplifies TLR7-mediated signaling during antiviral responses and autoimmunity. Nature immunology, 16(5), 495-504.
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