Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

AEBP2 (D7C6X) Rabbit mAb #14129

No. Size Price
14129S 100 µl ( 10 western blots ) ¥3,100.00 现货查询 购买询价
14129 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Monkey, Endogenous 70, 28 Rabbit IgG
IP 1:50
ChIP 1:25

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, ChIP=Chromatin IP,

Homology

Species predicted to react based on 100% sequence homology: Hamster, Chicken, Bovine,

Specificity / Sensitivity

AEBP2 (D7C6X) Rabbit mAb recognizes endogenous levels of total AEBP2 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu345 of human AEBP2 protein.

Western Blotting

Western Blotting

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

IP

IP

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.

Western Blotting

Western Blotting

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.

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 NCCIT cells and either 20 µl of AEBP2 (D7C6X) 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.

Background

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 PRC2 (EZH2-EED) complex is recruited to genes by DNA-binding transcription factors and methylates histone H3 on Lys27. Methylation of Lys27 facilitates the recruitment of the PRC1 complex, which ubiquitinylates histone H2A on Lys119 (5). Suppressor of Zeste 12 (SUZ12) is an obligate component of the PRC2 complex, which together with EZH2 and EED is absolutely required for histone methyltransferase activity of the protein complex (6).

The zinc finger AE binding protein 2 (AEBP2) is another integral component of the PRC2 complex. Addition of AEBP2 to the PRC2 core complex (EZH2-EED-SUZ12) enhances histone H3 Lys27 methyltransferase activity on nucleosomal substrates in vitro, which may be mediated in part by three AEBP2 DNA-binding zinc finger domains (5,7). AEBP2-mediated enhancement of enzymatic activity is greater on nucleosomal substrates that contain mono-ubiquitinated histone H2A Lys119, which suggests that AEBP2 may target PRC2 complexes in vivo through binding to DNA and mono-ubiquitinated histone H2A Lys119 (8).

  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. Wang, H. et al. (2004) Nature 431, 873-8.
  7. Kim, H. et al. (2009) Nucleic Acids Res 37, 2940-50.
  8. Kalb, R. et al. (2014) Nat Struct Mol Biol 21, 569-71.

Application References

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Protocols

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

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

XP is a registered trademark of Cell Signaling Technology, Inc.

SignalSilence is a registered trademark of Cell Signaling Technology, Inc.

SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.

Tween is a registered trademark of ICI Americas, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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