Revision 4

#3737Store 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, IF-IC, FC-FP, ChIP, ChIP-seq, C&R, eCLIP

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

83

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q15022

Entrez-Gene Id:

23512

Product Information

Product Usage Information

For optimal ChIP and ChIP-seq 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.

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Immunofluorescence (Immunocytochemistry) 1:800
Flow Cytometry (Fixed/Permeabilized) 1:100
Chromatin IP 1:100
Chromatin IP-seq 1:100
CUT&RUN 1:100
eCLIP 1:200
For more information about the RBP-eCLIP service please visit Eclipsebio.

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

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

Species Reactivity:

Human, Mouse, Rat, Monkey

Species predicted to react based on 100% sequence homology

Pig, Horse

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro257 of human Suz12 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. 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.

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 IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized) ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq C&R: CUT&RUN eCLIP: eCLIP

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.
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 4
#3737

SUZ12 (D39F6) XP® Rabbit mAb

Western Blotting Image 1: SUZ12 (D39F6) XP® Rabbit mAb Expand Image
使用 SUZ12 (D39F6) XP® Rabbit mAb 对不同细胞系的提取物进行蛋白质印迹分析。
No image available
Immunofluorescence Image 1: SUZ12 (D39F6) XP® Rabbit mAb Expand Image
使用 SUZ12 (D39F6) XP® Rabbit mAb(绿色)对 HeLa 细胞(左图)和 NTERA2 细胞(右图)进行共聚焦免疫荧光分析。肌动蛋白纤丝用 DY-554 Phalloidin 进行标记(红色)。
Immunofluorescence Image 2: SUZ12 (D39F6) XP® Rabbit mAb Expand Image
使用 SUZ12 (D39F6) XP® Rabbit mAb(绿色)对在小鼠胚胎成纤维细胞 (MEF) 饲养细胞中生长的小鼠胚胎干细胞进行共聚焦免疫荧光分析。肌动蛋白纤丝用 DY-554 Phalloidin 进行标记(红色)。
Flow Cytometry Image 1: SUZ12 (D39F6) XP® Rabbit mAb Expand Image
以浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)作为对照,使用 SUZ12 (D39F6) XP® Rabbit mAb(实线)对 Hela 细胞进行流式细胞分析。Anti-rabbit IgG (H+L)、F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4414 作为二抗。
Chromatin Immunoprecipitation Image 1: SUZ12 (D39F6) XP® Rabbit mAb Expand Image
使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003 对 NCCIT 细胞的交联染色质与 SUZ12 (D39F6) XP® Rabbit mAb 或 RING1B (D22F2) XP® Rabbit mAb 进行染色质免疫沉淀。使用 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。已知 SUZ12 和 RING1B 在染色质上互相结合。结果图显示 HOXD 基因上的 SUZ12 和 RING1B 结合,HOXD 基因是 SUZ12 和 RING1B 的已知靶标基因(参见包含 ChIP-qPCR 数据的其它结果图)。欲知其他 ChIP-seq 情况,请下载产品说明书。
Chromatin Immunoprecipitation Image 2: SUZ12 (D39F6) XP® Rabbit mAb Expand Image
使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003 对 NCCIT 细胞的交联染色质与 SUZ12 (D39F6) XP® Rabbit mAb 或 RING1B (D22F2) XP® Rabbit mAb 进行染色质免疫沉淀。使用 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。已知 SUZ12 和 RING1B 在染色质上互相结合。结果图显示 HOXA 基因上的 SUZ12 和 RING1B(上图)与 SUZ12 的已知靶标基因 HOXD 基因的结果(参见包含 ChIP-qPCR 数据的其它结果图)。
Chromatin Immunoprecipitation Image 3: SUZ12 (D39F6) XP® Rabbit mAb Expand Image
使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003 对 NCCIT 细胞的交联染色质与 SUZ12 (D39F6) XP® Rabbit mAb 或 Normal Rabbit IgG #2729 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。每份样品的免疫沉淀 DNA 含量使用与染色质输入总量(相当于1)相对应的信号表示。
CUT and RUN Image 1: SUZ12 (D39F6) XP® Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652,通过 NCCIT 细胞和 SUZ12 (D39F6) XP® Rabbit mAb 进行 CUT&RUN。使用 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。结果图显示在 SUZ12 的已知靶基因 HoxA1 处结合(参见包含 CUT&RUN-qPCR 数据的其他结果图)。
CUT and RUN Image 2: SUZ12 (D39F6) XP® Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652,通过 NCCIT 细胞和 SUZ12 (D39F6) XP® Rabbit mAb 进行 CUT&RUN。使用 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。结果图显示在 SUZ12 的已知靶基因 HoxD(上图)和 HoxA(下图)处结合(参见包含 CUT&RUN-qPCR 数据的其他结果图)。
CUT and RUN Image 3: SUZ12 (D39F6) XP® Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652,通过€‚ NCCIT 细胞和 SUZ12 (D39F6) XP® Rabbit mAb 或 Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362 进行 CUT&RUN。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。
eCLIP Image 1: SUZ12 (D39F6) XP® Rabbit mAb Expand Image
Enhanced cross-linking and immunoprecipitation (eCLIP) was performed with RNA from K-562 cells and SUZ12 (D39F6) XP® Rabbit mAb using a protocol based on the RBP-eCLIP method from Eclipsebio. 该图显示在 NOTCH1 转录内的结合。数据由 Gene Yeo 博士的实验室友情提供,并经许可使用。