Revision 5

#25069Store 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, C&T

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

70

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q14781

Entrez-Gene Id:

84733

Product Information

Product Usage Information

For optimal ChIP results, use 5 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.

The CUT&Tag dilution was determined using CUT&Tag Assay Kit #77552.

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Chromatin IP 1:100
Chromatin IP-seq 1:100
CUT&RUN 1:50
CUT&Tag 1:50

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

CBX2 (E3N6A) Rabbit mAb recognizes endogenous levels of total CBX2 protein.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala100 of human CBX2 protein.

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. This histone methyl-transferase activity requires the Ezh2, Eed, and Suz12 subunits of the complex (5). Histone H3 methylation at Lys27 facilitates the recruitment of the second complex, PRC1, which ubiquitinylates histone H2A on Lys119 (6). Chromobox protein homolog 2 (CBX2) is a core component of the PRC1 complex, which binds to the H3K27me3 mark and recruits PRC1 for transcriptional repression (7). CBX2 is frequently overexpressed in various cancers, including breast, ovarian, and AR-independent prostate cancer, where it is associated with poor prognosis (8-14). Furthermore, elevated CBX2 expression levels correlate with larger tumor size, increased lymph node metastasis, and positive human epidermal growth factor receptor 2 (HER-2) status in breast cancer (8). Mutations in CBX2 have also been attributed to 46,XY disorders of sex development (DSD), at least partially due to misregulation of the NR5A1/SF1 gene (15,16).

  1. Boyer, L.A. et al. (2006) Nature 441, 349-53.
  2. Lee, T.I. et al. (2006) Cell 125, 301-13.
  3. Park, I.K. et al. (2003) Nature 423, 302-5.
  4. Molofsky, A.V. et al. (2003) Nature 425, 962-7.
  5. Tavares, L. et al. (2012) Cell 148, 664-78.
  6. Gao, Z. et al. (2012) Mol Cell 45, 344-56.
  7. Kaustov, L. et al. (2011) J Biol Chem 286, 521-9.
  8. Chen, W.Y. et al. (2017) Oncol Lett 13, 1149-1156.
  9. Liang, Y.K. et al. (2017) Oncotarget 8, 92375-92387.
  10. Piqué, D.G. et al. (2019) Br J Cancer 120, 746-753.
  11. Zheng, S. et al. (2019) Am J Transl Res 11, 1668-1682.
  12. Iqbal, M.A. et al. (2021) Mol Oncol 15, 1450-1465.
  13. Wheeler, L.J. et al. (2018) Oncogenesis 7, 92.
  14. Clermont, P.L. et al. (2016) Clin Epigenetics 8, 16.
  15. Biason-Lauber, A. et al. (2009) Am J Hum Genet 84, 658-63.
  16. Sproll, P. et al. (2019) Sci Rep 9, 16552.

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 C&T: CUT&Tag

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.
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
仅供研究使用。不得用于诊断流程。
Revision 5
#25069

CBX2 (E3N6A) Rabbit mAb

Western Blotting Image 1: CBX2 (E3N6A) Rabbit mAb Expand Image
使用 CBX2 (E3N6A) Rabbit mAb（上图）或 β-Actin (D6A8) Rabbit mAb #8457（下图）对不同细胞系的提取物进行蛋白质印迹法分析。AsPC-1 细胞中 CBX2 蛋白的低表达与预测的表达模式一致。
Western Blotting Image 2: CBX2 (E3N6A) Rabbit mAb Expand Image
使用 CBX2 (E3N6A) Rabbit mAb 对不同细胞系的提取物进行蛋白质印迹法分析。
Immunoprecipitation Image 1: CBX2 (E3N6A) Rabbit mAb Expand Image
对 293T 细胞提取物的 CBX2 蛋白进行免疫沉淀分析。泳道 1 为 10% input，泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900，泳道 3 为 CBX2 (E3N6A) Rabbit mAb。使用 CBX2 (E3N6A) Rabbit mAb 进行蛋白质印迹法分析。Anti-rabbit IgG, HRP-linked Antibody #7074 用作二抗。
Chromatin Immunoprecipitation Image 1: CBX2 (E3N6A) Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005 对 NCCIT 细胞的交联染色质 和 CBX2 (E3N6A) Rabbit mAb, CBX2 Antibody #18687 或 RING1B (D22F2) XP® Rabbit mAb #5694 进行染色质免疫沉淀分析。使用 DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。该图显示 CBX2 的已知靶标基因 HoxA1 内的结合（参见包含 ChIP-qPCR 数据的其他结果图）。
Chromatin Immunoprecipitation Image 2: CBX2 (E3N6A) Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005 对 NCCIT 细胞的交联染色质 和 CBX2 (E3N6A) Rabbit mAb, CBX2 Antibody #18687 或 RING1B (D22F2) XP® Rabbit mAb #5694 进行染色质免疫沉淀分析。使用 DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。这些图显示 CBX2 的已知靶标基因 HoxA（上图）和 HoxD（下图）内的结合（参见包含 ChIP-qPCR 数据的其他结果图）。
Chromatin Immunoprecipitation Image 3: CBX2 (E3N6A) Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005 对 NCCIT 细胞的交联染色质与 CBX2 (E3N6A) Rabbit mAb 或 Normal Rabbit IgG #2729 进行染色质免疫沉淀分析。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: CBX2 (E3N6A) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652 对 NCCIT 细胞和 CBX2 (E3N6A) Rabbit mAb 进行 CUT&RUN 检测。使用 DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。该图显示 CBX2 的已知靶标基因 TAL1 内的结合（参见包含 CUT&RUN-qPCR 数据的其他结果图）。
CUT and RUN Image 2: CBX2 (E3N6A) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652 对 NCCIT 细胞和 CBX2 (E3N6A) Rabbit mAb 进行 CUT&RUN 检测。使用 DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。该图显示染色体 1（上图）内的结合，包括 CBX2 的已知靶标基因 TAL1（下图）（参见包含 CUT&RUN-qPCR 数据的其他结果图）。
CUT and RUN Image 3: CBX2 (E3N6A) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652 对 NCCIT 细胞和 CBX2 (E3N6A) Rabbit mAb 或 Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362 进行 CUT&RUN 检测。使用人 TAL1 启动子引物、人 FOXP2 启动子引物和人 PRSS1 上游引物，通过实时 PCR 对富集的 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 Tag Image 1: CBX2 (E3N6A) Rabbit mAb Expand Image
CUT&Tag was performed with NCCIT cells and CBX2 (E3N6A) Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figure shows binding across HOXA1, a known target gene of CBX2 (see our ChIP-qPCR figure).
CUT and Tag Image 2: CBX2 (E3N6A) Rabbit mAb Expand Image
CUT&Tag was performed with NCCIT cells and CBX2 (E3N6A) Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figures show binding across the HOXA gene cluser (upper), and the HOXD gene cluster (lower), which are known target genes of CBX2 (see our ChIP-qPCR figure).
图像库
更多了解我们如何获得图像
Orders: 877-616-CELL (2355)•[email protected]• Support: 877-678-TECH (8324)•[email protected]• Web: cellsignal.com
仅供研究使用。不得用于诊断流程。