The anti-E-Cadherin (Cellsignaling, 1:200) and anti-FOXP3 (Abcam, 1:100) were used as the primary antibodies. The cells were incubated with Alexa Fluor® 594 dye conjugated secondary antibody (ThermoFisher). The nucleus was stained by DAPI (ThermoFisher).
Alexa fluor 594 dye conjugated secondary antibody
Manufactured by Thermo Fisher Scientific
Sourced in United States
Alexa Fluor 594 dye-conjugated secondary antibody is a fluorescent labeling reagent. It is used to detect and visualize target proteins in immunoassays and other biological applications.
Automatically generated - may contain errors
Lab products found in correlation
Fluorescence mounting medium, by Agilent Technologies (2 mentions)
Fluorescence microscope, by Olympus (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Anti e cadherin, by Cell Signaling Technology (1 mentions)
Anti foxp3, by Abcam (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Hoechst 33342, by Merck Group (1 mentions)
3 protocols using alexa fluor 594 dye conjugated secondary antibody
1
Immunofluorescence Staining of E-Cadherin and FOXP3
2
EBV-Induced Cellular DNA Damage Response
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CNE1 cells with a cell count of 5×105 were inoculated in six-well plate preset with sterilized coverslips. After growing overnight, 100 MOI EBVs were used to infect the cells for 24 hours. Then the coverslips were taken out and the cells on them were fixed with 4% paraformaldehyde for 15 minutes at room temperature, after which the cells were permeabilized with 0.2% Triton X-100 for 5 minutes, followed by 30 minutes of blocking in 4% FBS. The cells were incubated with green fluorescence antibodies against γ-H2AX (1:200; Signalway Antibody, College Park, MD, USA) for 2 hours and then with an Alexa Fluor 594 dye-conjugated secondary antibody (1:500; Thermo Fisher Scientific) for 1 hour before counterstaining of the nuclei with DAPI (1:1,000; Sigma-Aldrich, St Louis, MO, USA) at room temperature. The slides were covered with fluorescence mounting medium (Dako, Glostrup, Denmark) and photographed under a fluorescence microscope (Olympus, Tokyo, Japan).
3
Immunofluorescence Staining for EBNA2 and γH2AX
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Cells were grown on sterilised coverslips and fixed with 4% paraformaldehyde (PFA) for 15 min at room temperature. The cells were then permeabilised with 0.2% Triton X-100 for 5 min followed by 30 min of blocking in 4% foetal bovine serum. The cells were incubated with primary antibodies against EBNA2 (1:200 dilution, Abcam, USA) or γH2AX (1:200 dilution, Signalway Antibody, USA) for 2 h and then with an Alexa Fluor 594 dye-conjugated secondary antibody (1:500 dilution, Thermo Fisher) for 1 h before counterstaining the nucleus with Hoechst 33342 (Sigma-Aldrich, USA) at room temperature. Finally, the slides were covered with fluorescence mounting medium (Dako) and photographed under a fluorescence microscope (Olympus, Tokyo, Japan).
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