Immunofluorescence[1]                     26.01.2003

PBS-Formalin

0.5ml    Paraformaldehyde 37%

9.5ml    PBS

PBS-FBS

400µl    fetal calf serum

20ml     PBS

5µl      CaCl₂ stock solution 1mol/l

Method

20min    Fixation in PBS-Formalin

3x10min  wash in PBS

5min     0.2% Triton X/PBS-FBS pH 7.3, 0°C

3x10min  wash in PBS-FBS

min 1h   primary antibody in PBS-FBS at RT

3x10min  wash in PBS-BSA

min 1h   sec. antibody 1:50 in PBS-FBS

         Phalloidin TRITC 0.1-1µM (Stock 100µM in DMSO)Sigma P1951
         DAPI 0.5µg/ml

2x10min  wash in PBS-FBS

 

Rat Bone Marrow Cells on stainless steel. 8h adhesion time. Green: Vinculin Red:    f-Actin Blue:   Nucleus