Western blot analysis of extracts from various cell lines using Ikaros (D10E5) Rabbit mAb (upper) or β-actin (D6A8) Rabbit mAb #8457 (lower). Western blot results confirm absence of detectable Ikaros protein in HeLa cells, in accordance with the immunofluorescence analysis.
Confocal immunofluorescent analysis of Jurkat cells (positive; left) and HeLa cells (negative; right) using Ikaros (D10E5) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).
Flow cytometric analysis of Ramos cells using Ikaros (D10E5) Rabbit mAb (blue) compared to concentration-matched Rabbit (DA1E) mAb IgG XP ® Isotype Control #3900 (red). 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 Jurkat cells and either 10 μl of Ikaros (D10E5) 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 HES1 promoter primers, SimpleChIP ® Human HES4 Promoter Primers #7273, 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.