The cells were seeded on cover slips and infected with C. trachomatis serovar L2 at MOI 1 for indicated time points. Before fixation with 4% PFA/Sucrose (Roth), the cells were washed with DPBS (Gibco). Fixed cells were permeabilized with 0.2% Triton X-100 (Sigma-Aldrich) in ×1 DPBS for 30 min, blocked with 2% FBS in ×1 DPBS for 45 min and incubated with primary antibodies for 1 hr at room temperature. Primary antibodies, chlamydial HSP60 (Santa Cruz, 1:500) and Phalloidin (Thermo Fisher Scientific), were diluted in 2% FBS in ×1 DPBS. Samples were washed and incubated with fluorescence dye conjugated secondary antibodies (Dianova) for 1 hr in the dark at room temperature. Cover slips were mounted onto microscopy slides using mowiol, slides were air-dried for at least 24 hr and examined using a LEICA DM2500 fluorescence microscope. The images were analysed with LAS AF program (Leica) and ImageJ software.
Fluorescence dye conjugated secondary antibodies
Manufactured by Dianova
Fluorescence dye conjugated secondary antibodies are laboratory reagents used in various immunoassay techniques. They are designed to bind to primary antibodies and emit fluorescent signals, allowing for the detection and visualization of target analytes in samples.
Automatically generated - may contain errors
Lab products found in correlation
Triton x 100, by Merck Group (2 mentions)
Dulbecco phosphate buffered saline (dpbs), by Thermo Fisher Scientific (2 mentions)
Chlamydial hsp60, by Santa Cruz Biotechnology (2 mentions)
Pfa sucrose, by Carl Roth (2 mentions)
Dulbecco phosphate buffered saline (dpbs), by Merck Group (1 mentions)
Las af program, by Leica (1 mentions)
Dm2500, by Leica (1 mentions)
2 protocols using fluorescence dye conjugated secondary antibodies
1
Immunofluorescence Assay for Chlamydia
2
Chlamydia trachomatis Immunofluorescence Assay
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The cells were seeded on cover slips and infected with Chlamydia trachomatis serovar L 2 at MOI 1 for indicated time points. Before fixation with 4% PFA/Sucrose (Roth), the cells were washed with DPBS (Gibco TM ). Fixed cells were permeabilized with 0.2% Triton-X-100 (Sigma-Aldrich) in 1x DPBS for 30 min, blocked with 2% FBS in 1x DPBS for 45 min and incubated with primary antibodies for 1 h at room temperature. Primary antibodies were diluted in 2% FBS in 1x DPBS; chlamydial HSP60 (Santa Cruz, 1:500) and Phalloidin (Thermo Fischer). Samples were washed and incubated with fluorescence dye conjugated secondary antibodies (Dianova) for 1 h in the dark at room temperature. Cover slips were mounted onto microscopy slides using mowiol, slides were air dried for at least 24 h and examined using a LEICA DM2500 fluorescence microscope. The images were analysed with LAS AF program and Image J software.
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