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9788
Polycomb Group Antibody Sampler Kit

Polycomb Group Antibody Sampler Kit #9788

Western Blotting Image 1

Western blot analysis of extracts from various cell lines using SUZ12 (D39F6) XP® Rabbit mAb.

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Chromatin IP-seq Image 2

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 NCCIT cells and either 5 µl of SUZ12 (D39F6) XP® Rabbit mAb or 10 μl of RING1B (D22F2) XP® Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. 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. SUZ12 and RING1B are known to associate with each other on chromatin. The figure shows binding of both SUZ12 and RING1B across HOXD genes, which are known target genes of both SUZ12 and RING1B (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.

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Western Blotting Image 3

Western blot analysis of extracts from MCF7, Neuro-2a, and COS-7 cell lines using Ezh2 (D2C9) XP® Rabbit mAb.

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Chromatin IP-seq Image 4

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 Hela cells and either 5 µl of Ezh2 (D2C9) XP® Rabbit mAb or 10 µl of Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared from 5ng enriched ChIP DNA for EZH2 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. EZH2 and H3K27me3 are known to associate with each other on chromatin. The figure shows binding of both EZH2 and H3K27me3 across the MYT1 gene. For additional ChIP-seq tracks, please download the product data sheet.

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Western Blotting Image 5

Western blot analysis of extracts from various cell lines using Ring1A (D2P4D) Rabbit mAb.

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Western Blotting Image 6

Western blot analysis of extracts from various cell lines using RING1B (D22F2) XP® Rabbit mAb.

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Chromatin IP-seq Image 7

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 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.

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Western Blotting Image 8

Western blot analysis of extracts from various cell lines using Bmi1 (D20B7) XP® Rabbit mAb.

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Chromatin IP-seq Image 9

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 Hela cells and either 5 µl of Ezh2 (D2C9) XP® Rabbit mAb #5246 or 10 µl of Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared from 5ng enriched ChIP DNA for EZH2 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. EZH2 and H3K27me3 are known to associate with each other on chromatin. The figure shows binding of both EZH2 and H3K27me3 across MYT1, a known target gene of H3K27me3 (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.

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Western Blotting Image 10

Western blot analysis of various cell lines using Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb.

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Western Blotting Image 11

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

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Chromatin IP Image 12

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 NCCIT cells and either 5 µl of SUZ12 (D39F6) 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).

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Flow Cytometry Image 13

Flow cytometric analysis of HeLa cells using SUZ12 (D39F6) XP® Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4414 was used as a secondary antibody.

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IHC-P (paraffin) Image 14

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Ezh2 (D2C9) XP® Rabbit mAb.

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Chromatin IP Image 15

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 NCCIT cells and either 5 µl of Ezh2 (D2C9) 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, which is equivalent to one.

Learn more about how we get our images
IF-IC Image 16

Confocal immunofluorescent analysis of HeLa cells using Ring1A (D2P4D) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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Chromatin IP Image 17

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 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).

Learn more about how we get our images
IHC-P (paraffin) Image 18

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Bmi1 (D20B7) XP® Rabbit mAb.

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Chromatin IP Image 19

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 10 µl of Tri-Methyl-Histone H3 (Lys27) (C36B11) 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, SimpleChIP® Human MyoD1 Exon 1 Primers #4490, and SimpleChIP® Human MYT-1 Exon 1 Primers #4493. 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.

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IHC-P (paraffin) Image 20

Immunohistochemical analysis of paraffin-embedded human lymphoma using Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb in the presence of non-methyl peptide (left) or K27 tri-methyl peptide (right).

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IF-IC Image 21

Confocal immunofluorescent analysis of HeLa cells (left) and NTERA2 cells (right) using SUZ12 (D39F6) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).

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IHC-P (paraffin) Image 22

Immunohistochemical analysis of paraffin-embedded human cervical carcinoma using Ezh2 (D2C9) XP® Rabbit mAb.

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IF-IC Image 23

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).

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IHC-P (paraffin) Image 24

Immunohistochemical analysis of paraffin-embedded human lymphoma using Bmi1 (D20B7) XP® Rabbit mAb.

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Flow Cytometry Image 25

Flow cytometric analysis of human whole blood cells using Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb (blue) and Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4414 was used as a secondary antibody. Analysis was performed on cells in the lymphocyte gate.

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IF-IC Image 26

Confocal immunofluorescent analysis of mouse embryonic stem cells growing on mouse embryonic fibroblast (MEF) feeder cells using SUZ12 (D39F6) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).

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IHC-P (paraffin) Image 27

Immunohistochemical analysis of paraffin-embedded human lymphoma using Ezh2 (D2C9) XP® Rabbit mAb.

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Flow Cytometry Image 28

Flow cytometric analysis of K562 cells using Bmi1 (D20B7) XP® Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

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IF-IC Image 29

Confocal immunofluorescent analysis of HeLa cells using Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).

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IHC-P (paraffin) Image 30

Immunohistochemical analysis of paraffin-embedded 4T1a mouse syngeneic tumor using Ezh2 (D2C9) XP® Rabbit mAb.

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IF-IC Image 31

Confocal immunofluorescent analysis of COS-7 cells using Bmi1 (D20B7) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).

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Flow Cytometry Image 32

Flow cytometric analysis of human peripheral blood mononuclear cells untreated (left) and treated (right) with anti-human CD3 (10ug/ml, coated plates) and anti-human CD28 (5ug/ml) for 3 days at 37ºC using EZH2 (D2C9) XP® Rabbit mAb and co-stained with an anti-human CD3 antibody. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor 488 Conjugate) #4412 was used as a secondary antibody.

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Chromatin IP Image 33

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 NCCIT cells and either 10 µl of Bmi1 (D20B7) 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).

Learn more about how we get our images
IF-IC Image 34

Confocal immunofluorescent analysis of HeLa cells using Ezh2 (D2C9) XP® Rabbit mAb (green) and S6 Ribosomal Protein (54D2) Mouse mAb #2317 (blue). Actin filaments were labeled with DY-554 phalloidin (red).

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IF-F Image 35

Confocal immunofluorescent analysis of mouse hippocampus (left) and cerebellum (right) using Ezh2 (D2C9) XP® Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
SUZ12 (D39F6) XP® Rabbit mAb 3737 20 µl
  • WB
  • IP
  • IF
  • F
  • ChIP
H M R Mk 83 Rabbit IgG
Ezh2 (D2C9) XP® Rabbit mAb 5246 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
  • ChIP
H M R Mk 98 Rabbit IgG
Ring1A (D2P4D) Rabbit mAb 13069 20 µl
  • WB
  • IP
  • IF
H R 54 Rabbit IgG
RING1B (D22F2) XP® Rabbit mAb 5694 20 µl
  • WB
  • IP
  • IF
  • ChIP
H M R Mk 41 Rabbit IgG
Bmi1 (D20B7) XP® Rabbit mAb 6964 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
  • ChIP
H Mk 41, 43 Rabbit IgG
Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb 9733 20 µl
  • WB
  • IHC
  • IF
  • F
  • ChIP
H M R Mk 17 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

The Polycomb Group Antibody Sampler Kit provides an economical means of evaluating total levels of Polycomb Group Proteins. The kit contains enough primary and secondary antibodies to perform two western mini-blot experiments.

Each antibody in the Polycomb Group Antibody Sampler Kit detects endogenous levels of target proteins.

Monoclonal antibody is produced by immunizing animals with synthetic peptides corresponding to residues surrounding Arg354 of human Ezh2, Pro316 of human Ring1A protein, the human SUZ12 protein, the carboxy terminus of human Bmi1 protein, the amino terminus of histone H3 in which Lys27 is tri-methylated, or with recombinant protein specific to the full-length human RING1B protein, respectively.

The polycomb group (PcG) proteins contribute to the maintenance of cell identity, stem cell self-renewal, cell-cycle regulation, and oncogenesis by maintaining the silenced state of genes that promote cell lineage specification, cell death, and cell-cycle arrest (1-4). PcG proteins exist in two complexes that cooperate to maintain long-term gene silencing through epigenetic chromatin modifications. The first complex, Eed-Ezh2, is recruited to genes by DNA-binding transcription factors and methylates histone H3 on Lys27. This histone methyltransferase activity requires the Ezh2, Eed, and Suz12 subunits of the complex (5). Methylation of Lys27 facilitates the recruitment of the second complex, PRC1, which ubiquitinates histone H2A on Lys119 (6). PRC1 is composed of Bmi1 and RING1A (also RING1 or RNF1), both of which act to enhance the E3 ubiquitin ligase activity of an additional catalytic subunit RING1B (also RING2 or RNF2) (7). PcG proteins play an important role in the regulation of cell proliferation and senescence through repression of the p16 INK4A and p19 ARF genes and are required for maintenance of adult hematopoietic and neural stem cells, as well as embryonic stem cells (3,4,8-10).

  1. Boyer, L.A. et al. (2006) Nature 441, 349-53.
  2. Lee, T.I. et al. (2006) Cell 125, 301-13.
  3. Park, I.K. et al. (2003) Nature 423, 302-5.
  4. Molofsky, A.V. et al. (2003) Nature 425, 962-7.
  5. Cao, R. and Zhang, Y. (2004) Mol Cell 15, 57-67.
  6. Wang, H. et al. (2004) Nature 431, 873-8.
  7. Cao, R. et al. (2005) Mol Cell 20, 845-54.
  8. Molofsky, A.V. et al. (2005) Genes Dev 19, 1432-7.
  9. Jacobs, J.J. et al. (1999) Nature 397, 164-8.
  10. Jacobs, J.J. et al. (1999) Genes Dev 13, 2678-90.
For Research Use Only. Not For Use In Diagnostic Procedures.

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To Purchase # 9788T

Product Number Size Price
9788T 1 Kit (6 x 20 µl) $409.00.0
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