Western blot analysis of extracts from various cell lines using Stat3 (D3Z2G) Rabbit mAb (upper), or β-Actin (D6A8) Rabbit mAb #8457 (lower). PC-3 cells are negative for Stat3.
Immunoprecipitation of Stat3 from HeLa cell extracts using Rabbit (DA1E) mAb IgG XP ® Isotype Control #3900 (lane 2) or Stat3 (D3Z2G) Rabbit mAb (lane 3). Lane 1 is 10% input.
Confocal immunofluorescent analysis of serum-starved HeLa cells, untreated (upper left) or treated with Human Interferon-α1 (hIFN-α1) #8927 (100 ng/ml, 30 min; upper right), and serum-starved PC-3 cells, untreated (lower right) or treated with Human Interferon-α1 (hIFN-α1) #8927 (100 ng/ml, 30 min; lower right), using Stat3 (D3Z2G) Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red).
Flow cytometric analysis of of PC-3 (blue) and HeLa (green) cells using Stat3 (D3Z2G) Rabbit mAb. Anti-rabbit IgG (H+L), F(ab') 2 Fragment (Alexa Fluor ® 488 Conjugate) #4412 was used as a secondary antibody.
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 10 6 Hep G2 cells starved overnight and treated with Human Interleukin-6 (hIL-6) #8904 (100 ng/ml) for 30 minutes, and either 10 μl of Stat3 (D3Z2G) 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 human IRF-1 promoter primers, SimpleChIP ® Human c-Fos Promoter Primers #4663, and SimpleChIP ® Human α Satellite Repeat Primers #4486. 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.