Experiments were carried out on cells seeded on cover slips. Cells were fixed with ice-cold methanol for 5 min at -20°C, washed with PBS and stained with DAPI, following which they were mounted onto glass slides using Prolong Gold. For nuclear translocation and co-localization assays, cells were fixed and permeabilized with 4% paraformaldehyde and 0.1% Triton-X. The cells were then blocked with 2.5% BSA and stained with the following antibodies: Rab7 (9367, Cell Signaling.), IRF3 (Cell Signaling, 4302), STING (AF6516), phospho S222 MVB12b (GenScript, Antibody production service) and corresponding secondary antibodies. Images were obtained on Zeiss LSM 710 confocal microscope using a 63× 1.4 oil-immersion objective and data were analyzed using ImageJ software. For click-it chemistry, EdC-labeled (T511307,Sigma) DNA of bacteria were detected in fixed cells according to manufacturer’s instructions (ThermoFisher, C10337) and AlexaFluor488 was detected under the microscope.
Sting
Manufactured by GenScript
STING is a laboratory equipment designed for the detection and measurement of the protein STING (Stimulator of Interferon Genes). It is a tool used in scientific research and investigations related to the STING signaling pathway and its role in cellular immune responses.
Automatically generated - may contain errors
2 protocols using sting
1
Imaging Nuclear Translocation and Colocalization
2
Imaging Nuclear Translocation and Colocalization
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Experiments were carried out on cells seeded on cover slips. Cells were fixed with ice-cold methanol for 5 min at -20°C, washed with PBS and stained with DAPI, following which they were mounted onto glass slides using Prolong Gold. For nuclear translocation and co-localization assays, cells were fixed and permeabilized with 4% paraformaldehyde and 0.1% Triton-X. The cells were then blocked with 2.5% BSA and stained with the following antibodies: Rab7 (9367, Cell Signaling.), IRF3 (Cell Signaling, 4302), STING (AF6516), phospho S222 MVB12b (GenScript, Antibody production service) and corresponding secondary antibodies. Images were obtained on Zeiss LSM 710 confocal microscope using a 63× 1.4 oil-immersion objective and data were analyzed using ImageJ software. For click-it chemistry, EdC-labeled (T511307,Sigma) DNA of bacteria were detected in fixed cells according to manufacturer’s instructions (ThermoFisher, C10337) and AlexaFluor488 was detected under the microscope.
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