Immunofluorescence staining

3.7% paraformaldehyde (100ml)
Heat 90ml of H2O to 60C
Add 3.7g of paraformaldehyde while stirring
Add 1 drop of 5M NaOH and stir till PFA dissolves completely
Add 10ml 10x PBS
Filter
Aliquot 1ml each
Keep at -20C
Warm till clear before use

0.2% Triton-X100 (10ml)
Triton-X100 20ul
1X PBS 9.98ml

Blocking buffer (10ml)
FBS 1ml1x
PBS 9ml

Seed 1.5 x 10^5 HCT116 cells in 12 well plate (1ml) with coverslips.
Label coverslip with letter to mark orientation.
Drug treatment
Transfer coverslips to parafilm in Petri dish.
Wash with PBS x2
Fix cells with 3.7% paraformaldehyde, 10min
Wash with PBS x2
Permeabilize cells with 0.2% Triton-X100, 10min
Wash with PBS x2
Block for 1h with blocking buffer
Incubate with primary antibodies (1:50-1:250) in blocking buffer, 1h
Wash with PBS x2Incubate with secondary antibodies (1:1000) in blocking buffer, 1h in the dark
Wash with PBS x2Incubate with DAPI (1:1000), 30min in dark
Wash with PBS x2
Mount coverslips on slides with FluroSave