Western blot analysis of extracts from HeLa cells, serum-starved overnight and then either untreated or stimulated for 4 hours with TPA (12-O-Tetradecanoylphorbol-13-Acetate) #4174, using Phospho-c-Fos (Ser32) (D82C12) XP ® Rabbit mAb (upper) and c-Fos (9F6) Rabbit mAb #2250 (lower). Antibody phospho-specificity is shown by treating lysates with λ-phosphatase.
Confocal immunofluorescent analysis of HeLa cells, serum-starved (left), TPA-treated (middle) or treated with TPA and λ-phosphatase (right), using Phospho-c-Fos (Ser32) (D82C12) XP ® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).
Flow cytometric analysis of HeLa cells, untreated (blue) or TPA-treated (green), using Phospho-c-Fos (Ser32) (D82C12) XP ® Rabbit mAb.
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 10 6 PC-12 cells starved overnight and treated with Human β-Nerve Gowth Factor (hβ-NGF) #5221 (50ng/ml) for 2h and either 10 μl of Phospho-c-Fos (Ser32) (D82C12) XP ® Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP ® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP ® Rat CCRN4L Promoter Primers #7983, rat DCLK1 promoter primers, and SimpleChIP ® Rat GAPDH Promoter Primers #7964. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.