Revision 6

#14129Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, 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 Information

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 Reactivity:

Human, Mouse, Rat, Monkey

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).

  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.

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

WB: 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 Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

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

Revision 6
#14129

AEBP2 (D7C6X) Rabbit mAb

Western Blotting Image 1: AEBP2 (D7C6X) Rabbit mAb Expand Image
使用 AEBP2 (D7C6X) Rabbit mAb 对不同细胞系提取物进行蛋白质印迹法分析。
Western Blotting Image 2: AEBP2 (D7C6X) Rabbit mAb Expand Image
使用 AEBP2 (D7C6X) Rabbit mAb (上)或 α-Actinin (D6F6) XP® Rabbit mAb #6487 (下),对细胞提取物进行蛋白质印迹法分析,这些细胞是使用 100 nM SignalSilence® Control siRNA(无偶联)#6568 (-)、SignalSilence® AEBP2 siRNA I #14711 (+) 或 SignalSilence® AEBP2 siRNA II #14713 (+) 转染的 HeLa 细胞。AEBP2 (D7C6X) Rabbit mAb 证实了 AEBP2 表达的沉默,而 α-Actinin (D6F6) XP® Rabbit mAb 则被用于上样对照。
Immunoprecipitation Image 1: AEBP2 (D7C6X) Rabbit mAb Expand Image
使用 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(泳道2)或 AEBP2 (D7C6X) Rabbit mAb(泳道3),对 NCCIT 细胞提取物 AEBP2 进行免疫沉淀。泳道 1 是 10% 输入对照。使用 AEBP2 (D7C6X) Rabbit mAb,进行蛋白质印迹法分析。
Chromatin Immunoprecipitation Image 1: AEBP2 (D7C6X) Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005 对 NCCIT 细胞中提取的交联染色质,在加入 JARID2 (D6M9X) Rabbit mAb #13594、CBX8 (D2O8C) Rabbit mAb #14696 或 AEBP2 (D7C6X) Rabbit mAb 后,进行染色质免疫沉淀。使用 DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。所有 JARID2、CBX8 和 AEBP2 已知可结合 PRC 复合体。结果图显示 JARID2、CBX8 和 AEBP2(JARID2、CBX8 和 AEBP2 的已知靶基因)在 HOXA 基因区域内的结合(参见含 ChIP-qPCR 数据的附加结果图)。
Chromatin Immunoprecipitation Image 2: AEBP2 (D7C6X) Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005 对 NCCIT 细胞中提取的交联染色质,在加入 JARID2 (D6M9X) Rabbit mAb #13594、CBX8 (D2O8C) Rabbit mAb #14696 或 AEBP2 (D7C6X) Rabbit mAb 后,进行染色质免疫沉淀。使用 DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。所有 JARID2、CBX8 和 AEBP2 已知可结合 PRC 复合体。本图显示整个 HOXA(上图)和 HOXD(下图)范围内 JARID2、CBX8 和 AEBP2(其为 JARID2、CBX8 和 AEBP2 的已知靶标基因)的结合作用(参见包含 ChIP-qPCR 数据的其他结果图)。
Chromatin Immunoprecipitation Image 3: AEBP2 (D7C6X) Rabbit mAb Expand Image
使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003,对 NCCIT 细胞中提取的交联染色质,在加入 AEBP2 (D7C6X) Rabbit mAb 或 Normal Rabbit IgG #2729 后,进行染色质免疫沉淀。使用 SimpleChIP® Human HoxA1 Intron 1 Primers #7707、SimpleChIP® Human HoxA2 Promoter Primers #5517 和 SimpleChIP® Human α Satellite Repeat Primers #4486 通过实时 PCR 对富集 DNA 进行定量分析。将每份样品中免疫沉淀的 DNA 的量表现为相对于所输入染色质总量(等于 1)的信号。
CUT and RUN Image 1: AEBP2 (D7C6X) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652,以 NCCIT 细胞和 AEBP2 (D7C6X) Rabbit mAb 进行 CUT&RUN。使用 DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。该图显示结合作用遍及 FZD5 基因(AEBP2 的已知靶标基因)(参见包含 CUT&RUN-qPCR 数据的其他结果图)。
CUT and RUN Image 2: AEBP2 (D7C6X) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652,以 NCCIT 细胞和 AEBP2 (D7C6X) Rabbit mAb 进行 CUT&RUN。使用 DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。该图显示结合作用遍及染色体 2(上图),包括 FZD5 基因(下图)(抗体简称的已知靶标基因)(参见包含 CUT&RUN-qPCR 数据的其他结果图)。
CUT and RUN Image 3: AEBP2 (D7C6X) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652,以NCCIT 细胞和处理 AEBP2 (D7C6X) Rabbit mAb 或 Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362进行 CUT&RUN。使用人 SOX2 启动子引物、人 FZD5 启动子引物和人 DTX2 启动子引物,通过实时 PCR 对富集的 DNA 定量。将每份样品中免疫沉淀的 DNA 的量表现为相对于所输入染色质总量(等于 1)的信号。