Western blot analysis of extracts from various cell lines using RING1B (D22F2) XP ® Rabbit mAb.
Confocal immunofluorescent analysis of HeLa cells using RING1B (D22F2) XP ® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5 ® #4084 (fluorescent DNA dye).
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 10 6 NCCIT cells and either 10 μl of RING1B (D22F2) XP ® Rabbit mAb or 10 µl of Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb #9733, using SimpleChIP ® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared from 5ng enriched ChIP DNA for RING1B ChIP-seq and 50ng enriched ChIP DNA for H3K27me3 ChIP-seq using NEBNext ® Ultra™ II DNA Library Prep Kit for Illumina ® , and sequenced on the Illumina NextSeq. RING1B and H3K27me3 are known to associate with each other on chromatin. The figure shows binding of both RING1B and H3K27me3 across HOXD genes, which are known target genes of RING1B (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 10 6 NCCIT cells and either 10 µl of RING1B (D22F2) 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 ® Human HoxA1 Intron 1 Primers #7707, SimpleChIP ® Human HoxA2 Promoter Primers #5517, 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 (equivalent to one).