Immunofluorescence[1] 26.01.2003

PBS-Formalin

0.5ml Paraformaldehyde 37%

9.5ml PBS

PBS-FBS

400l fetal calf serum

20ml PBS

5l 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, 0C

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 0.1-1M (Stock 100M in DMSO)Sigma P1951
DAPI 0.5g/ml

2x10min wash in PBS-FBS

 

Rat Bone Marrow Cells on stainless steel. 8h adhesion time.

Green: Vinculin

Red: f-Actin
Blue: Nucleus

 

 

 



[1] D.L. Spector and H.C. Smith, Redistribution of U-snRNPs during mitosis. Exp Cell Res, 1986. 163: 87-94.