Cells were analyzed as described previously.15 (link) Briefly, cells were fixed in 2% paraformaldehyde and permeabilized for 5 min in phosphate-buffered saline with 0.1% Triton-X-100. The primary antibody anti-γ-H2AX 3F2 antibody (1:200; Abcam, Cambridge, Great Britain) was used. Alexa 488-conjugated donkey anti-mouse (1:400; Invitrogen, Carlsbad, CA, USA) or Alexa 546-conjugated goat anti-mouse (1:200; Invitrogen) were used as secondary antibodies. For γ-tubulin staining, we used the ab11317 antibody (Abcam) at a 1:400 dilution and Alexa 546-conjugated goat anti-mouse antibody as the secondary antibody (1:200; Invitrogen). DNA was counterstained with 50 ng/ml DAPI (4,6-diamidino-2-phenylindole; Invitrogen). For γ-H2AX and Rad51 repair kinetics, cells were irradiated with 4 Gy (γ-irradiation) and incubated for 2, 4, 8 and 24 h at 37 °C followed by fixation and staining as described above.
Alexa 546 conjugated goat anti mouse antibody
Manufactured by Thermo Fisher Scientific
Alexa 546-conjugated goat anti-mouse antibody is a secondary antibody used for fluorescent labeling and detection of mouse primary antibodies in various immunoassays and imaging applications. The antibody is conjugated to the Alexa Fluor 546 dye, which has an excitation/emission spectrum suitable for fluorescence detection.
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Lab products found in correlation
Lsm 780, by Zeiss (2 mentions)
Alexa 488 conjugated donkey anti mouse, by Thermo Fisher Scientific (1 mentions)
Alexa 546 conjugated goat anti mouse, by Thermo Fisher Scientific (1 mentions)
Dapi 4 6 diamidino 2 phenylindole, by Thermo Fisher Scientific (1 mentions)
Anti histone h3 antibody, by Abcam (1 mentions)
Alexa 488 conjugated goat anti rabbit antibody, by Thermo Fisher Scientific (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
2 protocols using alexa 546 conjugated goat anti mouse antibody
1
Immunofluorescence Assay for DNA Damage
2
Immunofluorescence Staining of Viral-Infected Cells
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Briefly, cells on the coverslips were infected with WT virus at an MOI of 10. Thereafter, cells were fixed with 4% paraformaldehyde for 15 min, permeabilized with 0.1% TritonX-100 for 20 min, blocked in the PBS solution containing 5% BSA for 1 h, and incubated with primary rabbit polyclonal anti-Histone H3 antibody (Abcam) and mouse monoclonal anti-Lamin Dm0 antibody (DHSB) overnight at 4°C at indicated time points. Afterwards, cells were incubated with secondary Alexa 488-conjugated goat anti-rabbit antibody and Alexa 546-conjugated goat anti-mouse antibody (Invitrogen). Finally, cells were incubated with 4’,6-diamidino-2-phenylindole (DAPI, Beyotime), and examined through ZEISS LSM 780 confocal scanning laser microscopy (CSLM).
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