Choosing an antibody that recognizes the correct target protein and does not show non-specific binding with other DNA-binding proteins is critical for a successful ChIP-seq experiment. CST now tests and validates recombinant rabbit monoclonal antibodies in ChIP-seq, because good performance in ChIP-qPCR does not necessarily mean an antibody will perform well for ChIP-seq, which requires more extensive capture of the target protein across a large number of gene loci. Our ChIP-seq approved antibodies are tested using the SimpleChIP Enzymatic Chromatin IP protocol followed by next-generation sequencing (NGS).
CST has compared many of our ChIP-seq antibodies to ChIP-seq approved antibodies from other companies. Click on any of the CST antibodies below to see how we perform against the competition.
ChIP was performed with cross-linked chromatin from 4 x 106 MV-4-11 cells and either 7 μg of BRD4 (E2A7X) Rabbit mAb #13440 or 7 μg of Competitor Polyclonal Antibody, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared from 5 ng enriched ChIP DNA using NEBNext® Ultra™ II DNA Library Prep Kit for Illumina® and sequenced on the Illumina NextSeq. The CST recombinant rabbit monoclonal antibody provides higher signal and lower background than the Competitor Polyclonal Antibody in both whole genome analysis (Figure A) and localized gene analysis (Figure B).
ChIP was performed with cross-linked chromatin from 4 x 106 HeLa cells and either 1 μg of CTCF (D31H2) XP® Rabbit mAb #3418 or 1 μg of Competitor Polyclonal Antibody, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared from 5 ng enriched ChIP DNA using NEBNext® Ultra™ II DNA Library Prep Kit for Illumina® and sequenced on the Illumina NextSeq. The CST recombinant rabbit monoclonal antibody provides higher signal and lower background than the Competitor Polyclonal Antibody in both whole genome analysis (Figure A) and localized gene analysis (Figure B).
ChIP was performed with cross-linked chromatin from 4 x 106 HeLa cells and either 2 μg of Ezh2 (D2C9) XP® Rabbit mAb #5246 or 2 μg of Competitor Polyclonal Antibody, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared from 5 ng enriched ChIP DNA using NEBNext® Ultra™ II DNA Library Prep Kit for Illumina® and sequenced on the Illumina NextSeq. The CST recombinant rabbit monoclonal antibody provides equivalent signal and background levels as Competitor Polyclonal Antibody, as shown in the localized gene analysis.
ChIP was performed with cross-linked chromatin from 4 x 106 HeLa cells and either 3 μg of ENCODE-approved Mono-Methyl-Histone H3 (Lys4) (D1A9) XP® Rabbit mAb #5326 or 3 μg of ENCODE-approved Competitor Polyclonal Antibody, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared from 50 ng enriched ChIP DNA using NEBNext® Ultra™ II DNA Library Prep Kit for Illumina® and sequenced on the Illumina NextSeq. The CST recombinant rabbit monoclonal antibody provides higher signal and lower background than the ENCODE-approved Competitor Polyclonal Antibody, as shown in the localized gene analysis.
ChIP was performed with cross-linked chromatin from 4 x 106 HeLa cells and either 0.56 μg of Phospho-Rpb1 CTD (Ser2) (E1Z3G) Rabbit mAb #13499, 0.56 μg of ENCODE-approved Competitor Clone 3E10, or 0.56 μg of Competitor Polyclonal Antibody, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared from 5 ng enriched ChIP DNA using NEBNext® Ultra™ II DNA Library Prep Kit for Illumina® and sequenced on the Illumina NextSeq. The CST recombinant rabbit monoclonal antibody provides higher signal-to-noise ratio than the ENCODE-approved Competitor Clone 3E10 or the Competitor Polyclonal Antibody in whole genome analysis (Figure A) and shows Rpb1 binding across its known target gene ZNF740 (Figure B).
ChIP was performed with cross-linked chromatin from 4 x 106 HeLa cells and either 0.35 μg of Phospho-Rpb1 CTD (Ser5) (D9N5I) Rabbit mAb #13523, 0.35 μg of Competitor Polyclonal Antibody 1, or 0.35μg of Competitor Polyclonal Antibody 2, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared from 5 ng enriched ChIP DNA using NEBNext® Ultra™ II DNA Library Prep Kit for Illumina® and sequenced on the Illumina NextSeq. The CST recombinant rabbit monoclonal antibody provides higher signal and lower background than either Competitor Polyclonal Antibody 1 or 2 in both whole genome analysis (Figure A) and localized gene analysis (Figure B).
ChIP was performed with cross-linked chromatin from 4 x 106 HeLa cells and either 2 μg of Rpb1 NTD (D8L4Y) Rabbit mAb #14958 or 2 μg of Competitor Polyclonal Antibody, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared from 5 ng enriched ChIP DNA using NEBNext® Ultra™ II DNA Library Prep Kit for Illumina® and sequenced on the Illumina NextSeq. The CST recombinant rabbit monoclonal antibody provides higher signal and lower background than the Competitor Polyclonal Antibody in both whole genome analysis (Figure A) and localized gene analysis (Figure B).
ChIP was performed with cross-linked chromatin from 4 x 106 HeLa cells and either 1.65 μg of Tri-Methyl-Histone H3 (Lys9) (D4W1U) Rabbit mAb #13969, 2 µg of ENCODE-approved Competitor Polyclonal Antibody 1, or 2 μg of ENCODE-approved Competitor Polyclonal Antibody 2, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared from 50 ng enriched ChIP DNA using NEBNext® Ultra™ II DNA Library Prep Kit for Illumina® and sequenced on the Illumina NextSeq. The CST recombinant rabbit monoclonal antibody provides equivalent signal and background levels as Competitor Polyclonal Antibody 1 or 2, as shown in the localized gene analysis.
ChIP was performed with cross-linked chromatin from 4 x 106 NCCIT cells and either 0.59 μg of Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb #9733 or 2 μg of ENCODE-approved Competitor Polyclonal Antibody, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared from 50 ng enriched ChIP DNA using NEBNext® Ultra™ II DNA Library Prep Kit for Illumina® and sequenced on the Illumina NextSeq. The CST recombinant rabbit monoclonal antibody provides higher signal and lower background than the ENCODE-approved Competitor Polyclonal Antibody, as shown in the localized gene analysis.
