HEK293 cells were fixed with 4% paraformaldehyde in PBS for 20 min, and then permeabilized with 0.1% Triton X-100 in PBS on ice for 10 min. After rinsing with PBS buffer for three times, blocking solution (1% BSA in PBS) was applied for 1 h and primary antibody against Flag (Sigma, F1804, 1:200) was added in blocking buffer at 4 °C overnight. After washing with PBS/0.1% Triton X-100 for five times, cells were incubated with DAPI and secondary antibodies conjugated with Cy3 fluorescent dyes (Beyotime, a0521, 1:500) for 1 h, washed with PBS/0.1% Triton X-100, and mounted in Fluoromount-G (Southern Biotech). Images were recorded on a ZEISS Exciter 5 microscope (ZEISS).
Exciter 5 microscope
Manufactured by Zeiss
Sourced in Germany
The ZEISS Exciter 5 is a high-performance research microscope designed for advanced imaging and analysis applications. It features a versatile optical system, precise motorized controls, and integrated digital imaging capabilities to support a wide range of scientific investigations.
Automatically generated - may contain errors
Lab products found in correlation
Fluoromount g, by Southern Biotech (4 mentions)
Cy3 fluorescent dyes, by Beyotime (3 mentions)
Anti lamp2, by Santa Cruz Biotechnology (1 mentions)
Anti rpl4, by Santa Cruz Biotechnology (1 mentions)
Anti calnexin, by Proteintech (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Anti cox 4, by Proteintech (1 mentions)
Anti vimentin, by Santa Cruz Biotechnology (1 mentions)
4 protocols using exciter 5 microscope
1
Immunofluorescence Staining of Flag-Tagged Proteins
2
Immunofluorescence Microscopy of HeLa Cells
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HeLa cells were fixed with 4% paraformaldehyde in PBS for 20 min, and then permeabilized with 0.1% Triton X-100 in PBS on ice for 10 min. After rinsing with PBS buffer for three times, blocking solution (1% bovine serum albumin in PBS) was applied for 1 h and primary antibodies against Flag (Sigma,1:200) and SET7/9 (Upstate,1:200) were added in blocking buffer at 4 °C overnight. After washing with PBS/0.1% Triton X-100 for five times, cells were incubated with DAPI (4',6-Diamidino-2-phenylindole) and secondary antibodies conjugated with fluorescent dyes for 1 h, washed with PBS/0.1% Triton X-100, and mounted in Fluoromount-G (Southern Biotech, Birmingham, AL, USA). Images were recorded on a ZEISS Exciter 5 microscope (ZEISS, Jena, Germany).
3
Immunofluorescence Staining of Intracellular Organelles
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Cells were washed with phosphate-buffered saline (PBS) buffer for three times and then fixed with 4% paraformaldehyde in PBS buffer for 15 min and then permeabilized with 0.1% Triton X-100 in PBS buffer on ice for 10 min. After rinsing with PBS buffer for three times, blocking solution (5% bovine serum albumin in PBS buffer) was applied for 30 min and the corresponding primary antibody (anti-Calnexin (Proteintech, 10427-2-AP; 1:200 dilution), anti-COXIV (Proteintech, 11242-1-AP; 1:100), anti- EEA1 (home-made; 1:200 dilution), anti-LAMP2 (Santa Cruz Biotechnology, sc-18822; 1:200 dilution), anti-RPL4 (Santa Cruz Biotechnology, sc-100838; 1:200 dilution), anti-Vimentin (Santa Cruz Biotechnology, sc-373717; 1:200 dilution), or anti-Golgi 97 (Santa Cruz Biotechnology, sc-59820; 1:200 dilution) was added in blocking buffer at 4°C overnight. After washing with PBS buffer for five times, cells were incubated with secondary antibodies conjugated with Cy3 fluorescent dyes (Beyotime, a0521; 1:200) for 1 hour at room temperature and washed with PBS buffer for three times. 4′,6-Diamidino-2-phenylindole (1 mg/ml; Sigma-Aldrich) was then added for 5 min. Cells were then washed with PBS buffer for three times and mounted in Fluoromount-G (Southern Biotech). Images were recorded on a ZEISS Exciter 5 microscope (ZEISS).
4
Immunofluorescence Staining of HEK293 Cells
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HEK293 cells were fixed with 4% paraformaldehyde in PBS for 20 min, and then permeabilized with 0.1% Triton X‐100 in PBS on ice for 10 min. After rinsing with PBS buffer for three times, blocking solution (1% BSA in PBS) was applied for 1 h and primary antibody against Flag (Sigma, F1804, 1:200) was added in blocking buffer at 4 °C overnight. After washing with PBS/0.1% Triton X‐100 for five times, cells were incubated with DAPI and secondary antibodies conjugated with Cy3 fluorescent dyes (Beyotime, a0521, 1:500) for 1 h, washed with PBS/0.1% Triton X‐100, and mounted in Fluoromount‐G (Southern Biotech). Images were recorded on a ZEISS Exciter 5 microscope (ZEISS).
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