Subcellular localisation of PDCD4 was assessed by immunocytochemical staining. ARIP cells were seeded at 3×10⁵ cells/well on sterile cover slips in six well plates and incubated overnight under standard culture conditions.

Following specific experimental conditions, media was removed and cells were washed with PBS and fixed with 3.7% formalin. Cells were then permeabilised with 0.1% triton X-100 in PBS followed by blocking in blocking buffer (10% Goat serum; 2% BSA; 0.2% Tween 20; 0.7% Glycerol in PBS) for 1 hour. Cells were incubated overnight with ab51495 at 1/100 dilution followed by ab6717 (green) at 1/80 dilution. The nuclear counterstain is DAPI (Blue).

Results are representative of three separate experiments and images were representative of six separate fields. PDCD4 localized and expressing At G0: Cytoplasmic and very low expression; At N12: Cytoplasmic and low expression; At N24: very high cytoplasmic and low nuclear expression; At H12: Cytoplasmic expression and At H24 very high cytoplasmic and low nuclear expression. Over-all PDCD4 was highly expressed in the cytoplasm, with very low expression under serum starved conditions

ab11320 (Rabbit polyclonal to gamma Tubulin - Centrosome Marker) at 1/400 dilution staining human neuroblastoma cells by ICC/IF. The cells were fixed with paraformaldehyde and blocked with serum and then incubated with the primary antibody for 16 hours. ab6717 was used as the secondary.

Ab6717 was used at dilution 1/200 with the primary antibody ab36823 in IHC-P. See the review on ab36823.

Ab6717 was used with the primary antibody ab66506 in Flow Cyt. See Abreview on ab66506.