Fixed cells were permeabilized with 0.1% triton X-100 solution in 1X PBS. Cells were then blocked using Super Block (Scytek) and subsequently incubated overnight at 4°C with primary antibodies diluted in Super Block. Cells were washed and incubated with Alexa-Fluor secondary antibodies for 1 hour at room temperature. All antibody details are included in Table S1 . Nuclei were labelled with either 4′,6-diamidino-2-phenylindole (DAPI, ThermoFisher) or Hoechst 333432 (ThermoFisher). Cover slips were mounted on slides using ProLong antifade reagent (Invitrogen). Images were acquired using a 63X oil lens on Zeiss LSM 710 (Zeiss) or a Leica SP5 (Leica Microsystems) confocal microscope. Line scan analyses were performed to assess co-localization using Image J V1.52a33 .
Hoechst 333432
Manufactured by Thermo Fisher Scientific
Hoechst 333432 is a fluorescent dye used in cell biology and molecular biology applications. It is a cell-permeable nucleic acid stain that binds to DNA. The dye exhibits blue fluorescence upon binding to double-stranded DNA.
Automatically generated - may contain errors
2 protocols using hoechst 333432
1
Immunocytochemistry of Fixed Cells
2
Immunofluorescence Microscopy of Fixed Cells
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Fixed cells were permeabilized with 0.1% triton X-100 solution in 1X PBS. Cells were then blocked using Super Block (Scytek) and subsequently incubated overnight at 4°C with primary antibodies diluted in Super Block. Cells were washed and incubated with Alexa-Fluor secondary antibodies for 1 hour at room temperature. All antibodies used in this study are shown in Table S1. Nuclei were labelled with either 4K,6-diamidino-2-phenylindole (DAPI, ThermoFisher) or Hoechst 333432 (ThermoFisher).
Cover slips were mounted on slides using ProLong antifade reagent (Invitrogen). Images were acquired using a 63X oil lens on Zeiss LSM 710 (Zeiss) or a Leica SP5 (Leica Microsystems) confocal microscope.
Line scan analyses were performed to assess co-localization using Image J V1.52a (24) .
Cover slips were mounted on slides using ProLong antifade reagent (Invitrogen). Images were acquired using a 63X oil lens on Zeiss LSM 710 (Zeiss) or a Leica SP5 (Leica Microsystems) confocal microscope.
Line scan analyses were performed to assess co-localization using Image J V1.52a (24) .
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