Revision 6

#14129

Store at -20C

AEBP2 (D7C6X) Rabbit mAb
CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

Web:

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, IP, ChIP, ChIP-seq, C&R

Reactivity:
H M R Mk

Sensitivity:
Endogenous

MW (kDa):
70, 28

Source/Isotype:
Rabbit IgG

UniProt ID:
#Q6ZN18

Entrez-Gene Id:
121536

Product Usage Information

For optimal ChIP and ChIP-seq results, use 20 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.
Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Chromatin IP 1:25
Chromatin IP-seq 1:25
CUT&RUN 1:25

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity/Sensitivity

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

Species predicted to react based on 100% sequence homology

Hamster, Chicken, Bovine

Source / Purification

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

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).

Background References

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

W: Western Blotting IP: Immunoprecipitation ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq C&R: CUT&RUN

Cross-Reactivity Key

H: Human M: Mouse R: Rat Mk: Monkey

Trademarks and Patents

Cell Signaling Technology is a 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.

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

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意，否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件，除非书 CST 的合法授书代表以书面形式书独接受， 否书均被拒书，并且无效。

专品专有“专供研究使用”的专专或专似的专专声明， 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的， 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专，且专用于研专用途。将专品用于专断、专防或治专目的， 或专专售（专独或作专专成）或其他商专目的而专专专品，均需要 CST 的专独专可。客专：(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品，或使用专品制造任何商专专品，(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专，或使用专品开专任何与 CST 的专品或服专专争的专品或服专， (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专，(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品 ， (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

#14129

AEBP2 (D7C6X) Rabbit mAb

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using AEBP2 (D7C6X) Rabbit mAb.
Western Blotting Image 1: AEBP2 (D7C6X) Rabbit mAb
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.
Western Blotting Image 2: AEBP2 (D7C6X) Rabbit mAb
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.
Immunoprecipitation Image 1: AEBP2 (D7C6X) Rabbit mAb
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

#14129

AEBP2 (D7C6X) Rabbit mAb

Cell Signaling Technology Logo
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 DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #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).
Chromatin Immunoprecipitation Image 1: AEBP2 (D7C6X) Rabbit mAb
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 DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #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 (upper) and HOXD (lower), known target genes of JARID2, CBX8, and AEBP2 (see additional figure containing ChIP-qPCR data).
Chromatin Immunoprecipitation Image 2: AEBP2 (D7C6X) Rabbit mAb
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.
Chromatin Immunoprecipitation Image 3: AEBP2 (D7C6X) Rabbit mAb
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

#14129

AEBP2 (D7C6X) Rabbit mAb

Cell Signaling Technology Logo
CUT&RUN was performed with NCCIT cells and AEBP2 (D7C6X) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA library was prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figure shows binding across FZD5 gene, a known target gene of AEBP2 (see additional figure containing CUT&RUN-qPCR data).
CUT & RUN Image 1: AEBP2 (D7C6X) Rabbit mAb
CUT&RUN was performed with NCCIT cells and AEBP2 (D7C6X) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA library was prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 2 (upper), including FZD5 gene (lower), a known target gene of antibody’s short name (see additional figure containing CUT&RUN-qPCR data).
CUT & RUN Image 2: AEBP2 (D7C6X) Rabbit mAb
CUT&RUN was performed with NCCIT cells and treatment and either AEBP2 (D7C6X) Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using human SOX2 promoter primers, human FZD5 promoter primers, and human DTX2 promoter primers. 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.
CUT & RUN Image 3: AEBP2 (D7C6X) Rabbit mAb
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.