Western blot analysis of extracts from various cell lines using HP1β (D2F2) XP ® Rabbit mAb.
Confocal immunofluorescent analysis of C2C12 cells using HP1β (D2F2) XP ® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 10 6 Hela cells and either 10 µl of HP1β (D2F2) XP ® Rabbit mAb or 10 μl of Tri-Methyl-Histone H3 (Lys9) (D4W1U) Rabbit mAb #13969, using SimpleChIP ® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared from 5ng enriched ChIP DNA for HP1β ChIP-seq and 50ng enriched ChIP DNA for H3K9me3 ChIP-seq using NEBNext ® Ultra™ II DNA Library Prep Kit for Illumina ® , and sequenced on the Illumina NextSeq. HP1β and H3K9me3 are known to associate with each other on chromatin. The figure shows binding of both HP1β and H3K9me3 across ZNF genes, known target genes of both HP1β and H3K9me3. For additional ChIP-seq tracks, please download the product data sheet.
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 10 6 mES cells and either 10 µl of HP1β (D2F2) XP® Rabbit mAb #8676 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 ® Mouse MEST Intron 1 Primers #12870, SimpleChIP ® Mouse MEST Promoter Primers #12928, mouse Intracisternal A-Particle (IAP) LTR promoter primers, and mouse primers specific to a non-genic, non-repetitive region. 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.