Write a paper on how Cells were seeded two days before the ICC procedure on the sterilized coverslips in 48-well plates. After 48 hours of seeding, cells were washed with 1X DPBS 3 times and fixed with 4% PFA (Sigma-Aldrich). Then cells were washed with 1X PBS 3 times for permeabilized with Triton (Sigma-Aldrich) or Saponin(Sigma-Aldrich) in DPBS at room temperature, followed by washing the cells with cold DPBS 3 times. and then the cells were incubated with 3% Bovine Serum Albumin (BSA, (Sigma-Aldrich) in DPBS for 20 minutes at room temperature followed by washing the cells with cold DPBS 3 times. Cells were then treated with the anti-CFTR primary antibody (1:200, CFF570) in 0.1% BSA-DPBS, incubating overnight at 4 degrees. The next day primary antibody was removed by washing the cells with cold DPBS 3 times. The goat anti-mouse secondary antibody (Alexa Fluor 568) was used at 1:800 in 0.1% BSA-DPBS for 2 hours at room temperature (in a dark environment). Then cells were washed with cold DPBS 3 times and a drop of DAPI (ab104139, Abcam), was used to mount the coverslips and stain the nucleus. Control samples were used in this procedure with DAPI only and with secondary antibody and DAPI only. Mount a coverslip on the samples with the drops and incubate for 10 minutes in a dark environment. Cells were visualized using a fluorescence microscope.
This is the protocol but i want as a graphical summary. please attached this to the expert
