Cell Signaling Technology

Product Pathways - Chromatin Regulation

SUZ12 (D39F6) XP™ Rabbit mAb #3737

This XP™ monoclonal antibody was developed using our eXceptional Monoclonal Technology (XMT™).

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IF-IC ChIP H M R Mk Endogenous 83 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)  ChIP=Chromatin IP
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey
Species cross-reactivity is determined by Western blot.

Protocols

Specificity / Sensitivity

SUZ12 (D39F6) XP™ Rabbit mAb detects endogenous levels of SUZ12 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the sequence of the human SUZ12 protein.

Western Blotting

Western Blotting

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

IF-IC

IF-IC

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).

IF-IC

IF-IC

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).


Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 5 μl of SUZ12 (D39F6) XP™ Rabbit mAb #3737 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 primer sets specific to MYT-1 (Promoter), MYT-1 (-1700) and the heterochromatic α Satellite repeat element (MYT-1 primer sequences from reference 8). 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.

Background

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. Methylation of Lys27 facilitates the recruitment of the second complex, PRC1, which ubiquitinylates histone H2A on Lys119 (5). Suppressor of Zeste 12 (SUZ12) is a component of the PRC2 complex, which together with Ezh2 and Eed is absolutely required for histone methyl-transferase activity (6). SUZ12 contains a C2H2 zinc finger domain similar to the zinc finger domains found in sequence-specific DNA binding proteins and may mediate the interaction between EZH2 and nucleosomes (6). SUZ12 is overexpressed in several human tumors, including tumors of the colon, breast and liver (7,8).

  1. Boyer, L.A. et al. (2006) Nature 441, 349-53.
  2. Lee, T.I. et al. (2006) Cell 125, 301-13.
  3. Cao, R. et al. (2002) Science 298, 1039-43.
  4. Müller, J. et al. (2002) Cell 111, 197-208.
  5. Wang, H. et al. (2004) Nature 431, 873-8.
  6. Cao, R. and Zhang, Y. (2004) Mol Cell 15, 57-67.
  7. Kirmizis, A. et al. (2003) Mol Cancer Ther 2, 113-21.
  8. Kirmizis, A. et al. (2004) Genes Dev 18, 1592-605.

Application References

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This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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