Western blot analysis of extracts from HeLa and C6 cells, untreated (-) or treated (+) with Trichostatin A (TSA) #9950 (1 μM, 18 hr), using Acetyl-Histone H4 (Lys12) (D2W6O) Rabbit mAb (upper) and Histone H4 (D2X4V) Rabbit mAb #13919 (lower).
Confocal immunofluorescent analysis of HeLa cells untreated (left) or treated (right) with Trichostatin A (TSA) #9950 (1 µM, 4 hr) using Acetyl-Histone H4 (Lys12) (D2W60) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red).
Flow cytometric analysis of HeLa cells, untreated (blue) or treated with SAHA (green) using Acetyl-Histone H4 (Lys12) (D2W6O) Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 fragment (Alexa Fluor ® 488 Conjugate) #4412 was used as secondary antibody.
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 10 6 HeLa cells and either 10 μl of Acetyl-Histone H4 (Lys12) (D2W6O) 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 ® Human GAPDH Exon 1 Primers #5516, SimpleChIP ® Human RPL30 Exon 3 Primers #7014, 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.