Target/Analyte:
DNA | F-Actin | 2 cell markers

Short Name DNA-ACTIN-IF
Description DNA imaged with DAPI, F-Actin imaged with Alexa Fluor 647 Phalloidin and two variable cell markers labeled with a pair of primary antibodies (rabbit and mouse) and the corresponding secondary antibodies (conjugated to AF488 and AF561 respectively)

Method/Kits with this Target/Analyte
Method/Kit Description
Dual label immunofluorescence with DAPI and phalloidin counterstain Fixed cells are permeabilized with triton X-100 and incubated in blocking solution (depending on the antibodies used), then stained with Alexa Fluor 647 phalloidin (concentration optimized per cell line) for 2 hrs. at room temperature, followed by overnight incubation at 4°C with a mixture of two primary antibodies (concentration optimized per cell line and target) and then by 1 hr. incubation at room temperature with a secondary antibody mixture (concentration optimized depending on target and sample). Finally cells are stained with DAPI (1 µg/ml) for 10 min at room temperature. Final processing depends on the nature of the sample, tissue sections, cells on coverslips, transwell membrane fragments, are mounted (mounting medium depending on the sample) on microscope slides. Cells grown on multiwell plates (including mimetas plates) are images directly after the final wash. Imaging performed by fluorescence microscopy (widefield or confocal)
Dual label immunofluorescence with DAPI and phalloidin counterstain Fixed cells are permeabilized with triton X-100 and incubated in blocking solution (depending on the antibodies used), then stained with Alexa Fluor 647 phalloidin (concentration optimized per cell line) for 2 hrs. at room temperature, followed by overnight incubation at 4°C with a mixture of two primary antibodies (concentration optimized per cell line and target) and then by 1 hr. incubation at room temperature with a secondary antibody mixture (concentration optimized depending on target and sample). Finally cells are stained with DAPI (1 µg/ml) for 10 min at room temperature. Final processing depends on the nature of the sample, tissue sections, cells on coverslips, transwell membrane fragments, are mounted (mounting medium depending on the sample) on microscope slides. Cells grown on multiwell plates (including mimetas plates) are images directly after the final wash. Imaging performed by fluorescence microscopy (widefield or confocal)

Studies Utilizing this Target/Analyte
Study Start Date Study Types Description