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62083
Polycomb Group 2 (PRC2) Antibody Sampler Kit
Primary Antibodies

Polycomb Group 2 (PRC2) Antibody Sampler Kit #62083

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Chromatin immunoprecipitations were performed with cross-linked chromatin from Hela cells and either Ezh2 (D2C9) XP® Rabbit mAb or Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. 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.

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

Western blot analysis of extracts from various cell lines using EZH1 (D7D5D) Rabbit mAb.

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

Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and either SUZ12 (D39F6) XP® Rabbit mAb or RING1B (D22F2) XP® Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. 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.

Western blot analysis of extracts from 293T cells, either transfected with nontargeting siRNA (293T siNT) or siRNA targeting EED (293T siEED), and F9 cells using EED Antibody (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).

Western blot analysis of extracts from F9 cells and NTERA-2 cells, untreated (-) or treated with retinoic acid (1 μM, 5 d; +) to induce differentiation, using JARID2 (D6M9X) Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). As expected, JARID2 protein levels decrease upon NTERA-2 cell differentiation.

Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and either JARID2 (D6M9X) Rabbit mAb, CBX8 (D2O8C) Rabbit mAb #14696, or AEBP2 (D7C6X) Rabbit mAb #14129, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. All of JARID2, CBX8, and AEBP2 are known to associate with PRC complexes. The figure shows binding of JARID2, CBX8, and AEBP2 across HOXD gene region, known target genes of JARID2, CBX8, and AEBP2 (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 NCCIT cells and either JARID2 (D6M9X) Rabbit mAb #13594, CBX8 (D2O8C) Rabbit mAb #14696, or AEBP2 (D7C6X) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. All of JARID2, CBX8, and AEBP2 are known to associate with PRC complexes. The figure shows binding of JARID2, CBX8, and AEBP2 across HOXA gene region, known target genes of JARID2, CBX8, and AEBP2 (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.

Western blot analysis of extracts from various cell lines using AEBP2 (D7C6X) Rabbit mAb.

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.

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

Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and either Ezh2 (D2C9) XP® Rabbit mAb or 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.

Immunoprecipitation of EZH1 from NCCIT cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is EZH1 (D7D5D) Rabbit mAb. Western blot analysis was performed using EZH1 (D7D5D) Rabbit mAb.

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.

Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and either SUZ12 (D39F6) XP® Rabbit mAb or 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).

Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and either JARID2 (D6M9X) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. 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.

Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and either AEBP2 (D7C6X) Rabbit mAb or 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.

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® AEBP2 siRNA I #14711 (+), or SignalSilence® AEBP2 siRNA II #14713 (+), using AEBP2 (D7C6X) Rabbit mAb (upper) or α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower). The AEBP2 (D7C6X) Rabbit mAb confirms silencing of AEBP2 expression, while the α-Actinin (D6F6) XP® Rabbit mAb is used as a loading control.

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

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).

Immunoprecipitation of AEBP2 from NCCIT cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or AEBP2 (D7C6X) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using AEBP2 (D7C6X) Rabbit mAb.

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

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).

Immunohistochemical analysis of paraffin-embedded 4T1 mouse syngeneic tumor using Ezh2 (D2C9) XP® Rabbit mAb.

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.

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).

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).

To Purchase # 62083T
Product # Size Price
62083T
1 Kit  (6 x 20 µl) N/A

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Ezh2 (D2C9) XP® Rabbit mAb 5246 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
  • ChIP
H M R Mk 98 Rabbit IgG
EZH1 (D7D5D) Rabbit mAb 42088 20 µl
  • WB
  • IP
H Mk 95 Rabbit IgG
SUZ12 (D39F6) XP® Rabbit mAb 3737 20 µl
  • WB
  • IP
  • IF
  • F
  • ChIP
H M R Mk 83 Rabbit IgG
EED Antibody 51673 20 µl
  • WB
H M R Mk 50-70 Rabbit 
JARID2 (D6M9X) Rabbit mAb 13594 20 µl
  • WB
  • IP
  • ChIP
H M 150 Rabbit IgG
AEBP2 (D7C6X) Rabbit mAb 14129 20 µl
  • WB
  • IP
  • ChIP
H M R Mk 70, 28 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product Description

The Polycomb Group 2 (PRC2) Antibody Sampler Kit provides an economical means of evaluating total levels of Polycomb Group 2 Proteins. The kit contains enough primary and secondary antibodies to perform two western blot experiments.

Specificity / Sensitivity

Each antibody in the Polycomb Group 2 (PRC2) Antibody Sampler Kit detects endogenous levels of target proteins.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Arg354 of human Ezh2, Pro257 of human SUZ12 protein, Leu23 of human EED, Asp1114 of human JARID2, Leu345 of human AEBP2, and the carboxy terminus of EZH1.

Background

The polycomb group (PcG) proteins are involved in maintaining the silenced state of multiple developmentally regulated genes and contribute to the maintenance of cell identity, cell cycle regulation, and oncogenesis (1-4). Enhancer of zest homolog 1 (Ezh1) and enhancer of zest homolog 2 (Ezh2) are members of this large protein family and are subunits of the polycomb repressor complex 2 (PRC2), also known to contain SUZ12, EED, JARID2, and AEBP2. Ezh1 and its paralog Ezh2 are mutually exclusive catalytic subunits of the PRC2 complex, which functions to mono-, di-, and tri-methylate Lys27 on histone H3, all marks that are associated with transcriptional repression. SUZ12 and EED proteins are also absolutely required for methyltransferase activity (5). JARID2 and AEBP2 are both accessory proteins that function to recruit the PRC2 complex to target genes and enhance methyltransferase activity by binding to DNA and histone proteins in nucleosomes (6-14).

  1. Boyer, L.A. et al. (2006) Nature 441, 349-53.
  2. Cao, R. et al. (2002) Science 298, 1039-43.
  3. Müller, J. et al. (2002) Cell 111, 197-208.
  4. Lee, T.I. et al. (2006) Cell 125, 301-13.
  5. Cao, R. and Zhang, Y. (2004) Mol Cell 15, 57-67.
  6. Peng, J.C. et al. (2009) Cell 139, 1290-302.
  7. Shen, X. et al. (2009) Cell 139, 1303-14.
  8. Pasini, D. et al. (2010) Nature 464, 306-10.
  9. Li, G. et al. (2010) Genes Dev 24, 368-80.
  10. Son, J. et al. (2013) Genes Dev 27, 2663-77.
  11. Kaneko, S. et al. (2014) Mol Cell 53, 290-300.
  12. Cao, R. and Zhang, Y. (2004) Mol Cell 15, 57-67.
  13. Kim, H. et al. (2009) Nucleic Acids Res 37, 2940-50.
  14. Kalb, R. et al. (2014) Nat Struct Mol Biol 21, 569-71.

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For Research Use Only. Not For Use In Diagnostic Procedures.

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