Immunofluorescence Visualization of Rab7 and TLR7

Immunofluorescence staining was performed as described previously [87 (link)] using anti-Rab7 (diluted 1:100, Cell Signaling Technology, Danvers, Massachusetts, USA), anti-TLR7 (diluted 1:500, Novus Biologicals, Littleton, Colorado, USA), and Alexa Fluor 488 goat anti-rabbit IgG (diluted 1:2000, Thermo Fisher Scientific) as primary and secondary antibodies, respectively. After DNA counterstaining with ProLong Gold Antifade Reagent with DAPI (Thermo Fisher Scientific), images were acquired using a Nikon Eclipse Ti-U confocal microscope (Melville, New York, USA) at the Bioimaging Facility of the Sidney Kimmel Cancer Center at TJU.

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Pawar K., Shigematsu M., Sharbati S, & Kirino Y. (2020). Infection-induced 5′-half molecules of tRNAHisGUG activate Toll-like receptor 7. PLoS Biology, 18(12), e3000982.

Publication 2020

anti-Rab7 anti-TLR7 Alexa Fluor 488 goat anti-rabbit IgG ProLong Gold Antifade Reagent with DAPI Nikon Eclipse Ti-U confocal microscope

Alexa fluor 488 Anti igg Antibodies Biologicals Cancer Confocal microscope Dapi Goat Gold Immunofluorescence Novus Rabbit

Corresponding Organization : Freie Universität Berlin

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Variable analysis

independent variables

Antibody dilutions: anti-Rab7 (1:100), anti-TLR7 (1:500), Alexa Fluor 488 goat anti-rabbit IgG (1:2000)

dependent variables

Localization and distribution of Rab7 and TLR7 proteins in cells

control variables

Immunofluorescence staining protocol
DNA counterstaining with ProLong Gold Antifade Reagent with DAPI
Imaging using Nikon Eclipse Ti-U confocal microscope at the Bioimaging Facility of the Sidney Kimmel Cancer Center at TJU

controls

No positive or negative controls were explicitly mentioned in the provided information.

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