IMMUNOFLUORESCENCE ON CELLS IN CULTURE (1)

(incubation at 22°C on rotating-plate for good mixing)

• Cells cultured on coverslips 1.2 mm (100.000 cells/well)
   
• Wash 3 x 5 min with PBS-Ca²⁺, 37°C
   
• Fixation in 100 % methanol for 10 min at -20°C
   
• Wash 2 x 1 min with PBS-Ca²⁺, 22°C
   
• Incubation: first antibody 60 min in PBS-Ca²⁺ with 0.1% albumin
   
• Wash 5 x 5 min in PBS-Ca²⁺ with 0.1% albumin
   
• Incubation: second antibody 60 min in PBS-Ca²⁺ with 0.1% albumin in dark
   
• Wash 5 x 5 min in PBS-Ca²⁺ with 0.1% albumin
   
• Mounting coverslip in Glycerol/PBS/DABCO