Immunofluorescence detection of CPAF in chlamydia-infected cells was carried out as described previously 6 7 . In brief, HeLa cell monolayer was infected with Chlamydia trachomatis L2 for 30 h. The monolayer, after it was fixed with paraformaldehyde (Sigma-Aldrich) and permeabilized with Saponin (Sigma-Aldrich), was costained with Hoechst 32258 (blue), anti-MOMP antibody MC22 (probed with an FITC-conjugated, mouse IgG3-specific secondary antibody), and anti-CPAFn antibody 54b (probed with a Cy3-conjugated, mouse IgG1-specific secondary antibody). Images were acquired individually for each stain in gray using a Cooker digital camera connected to an AX70 Olympus microscope, and the single-color images were merged in frame into the triple-color image using the software Image Pro.
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Immunofluorescence Detection of CPAF in Chlamydia
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Anti antibody
Antibody
Antibody igg1
Cells
Chlamydia
Chlamydia trachomatis
Cooker
Cpaf
Fitc
Frame
Hela cell
Hoechst 32258
Igg3
Immunofluorescence
Microscope
Mouse
Paraformaldehyde
Saponin
Stain
Corresponding Organization :
Other organizations : The University of Texas Health Science Center at San Antonio, University of Manitoba
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Variable analysis
independent variables
- Chlamydia trachomatis L2 infection
- Immunofluorescence detection of CPAF in Chlamydia-infected cells
- HeLa cell monolayer
- Fixation with paraformaldehyde
- Permeabilization with Saponin
- Staining with Hoechst 32258 (blue), anti-MOMP antibody MC22 (probed with an FITC-conjugated, mouse IgG3-specific secondary antibody), and anti-CPAFn antibody 54b (probed with a Cy3-conjugated, mouse IgG1-specific secondary antibody)
- No positive or negative controls were explicitly mentioned.
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