Revision 4

#27945Store 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

REACTIVITY:

H Mk

SENSITIVITY:

Endogenous

MW (kDa):

180

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q9ULU4

Entrez-Gene Id:

23613

Product Information

Product Usage Information

For optimal ChIP and ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 × 10^6 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Chromatin IP 1:50
Chromatin IP-seq 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

ZMYND8 (E2Z4U) Rabbit mAb recognizes endogenous levels of total ZMYND8 protein.

Species Reactivity:

Human, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human ZMYND8 protein.

Background

Zinc finger MYND domain-containing protein 8 (ZMYND8), also referred to as receptor for activated C-kinase 7 (Rack7) and protein kinase C-binding protein 1 (PRKCBP1), is a DNA damage response protein and a transcriptional regulator that is a close homolog of ZMYND11 (BS69) (1). ZMYND8 binds to H3K36me2 and H4K16ac, two histone marks associated with active transcription (2). This protein is targeted to sites of DNA damage within actively transcribed genes, and recruits the H3K4me3-specific histone demethylase KDM5A/JARID1A and nucleosome remodeling and histone deacetylation (NuRD) complex (1-3). Together, these protein complexes mediate transcriptional repression and allow for subsequent double-strand break repair via homologous recombination. ZMYND8 contains a bromodomain and a PWWP domain near its N-terminus, and a MYND domain towards the C-terminus, the latter of which mediates interaction with the NuRD complex (1). ZMYND8 also functions to recruit the H3K4me3-specific histone demethylase KDM5C/JARID1C to enhancer and super-enhancer regions, and functions as a negative regulator of gene expression (4). ZMYND8 and JARID1C are both putative tumor suppressor proteins, and knockdown of either of these proteins leads to derepression of S100 oncogenes (1). ZMYND8 expression is altered in breast and cervical cancer (4,5), and has been found to be translocated with RELA in at least one patient with acute erythroid leukemia (6). Knockdown of ZMYND8 expression in breast cancer cell lines increases anchorage-independent cell growth, cell migration and invasion, and tumor growth in mouse xenograft models (4).

  1. Gong, F. et al. (2015) Genes Dev 29, 197-211.
  2. Adhikary, S. et al. (2016) J Biol Chem 291, 2664-81.
  3. Gong, F. et al. (2017) J Cell Biol 216, 1959-74.
  4. Shen, H. et al. (2016) Cell 165, 331-42.
  5. Bierkens, M. et al. (2013) Genes Chromosomes Cancer 52, 56-68.
  6. Panagopoulos, I. et al. (2013) PLoS One 8, e63663.

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 nonfat dry milk, 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

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

Revision 4
#27945

ZMYND8 (E2Z4U) Rabbit mAb

Western Blotting Image 1: ZMYND8 (E2Z4U) Rabbit mAb Expand Image
使用 ZMYND8 (E2Z4U) Rabbit mAb 对 HDLM-2、HeLa 和 HT-1080 细胞的提取物进行蛋白印迹分析。
Immunoprecipitation Image 1: ZMYND8 (E2Z4U) Rabbit mAb Expand Image
对 HT-1080 细胞提取物 ZMYND8 蛋白进行免疫沉淀。泳道 1 为 10% 输入,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 ZMYND8 (E2Z4U) Rabbit mAb。使用 ZMYND8 (E2Z4U) Rabbit mAb 进行蛋白印迹分析。
Chromatin Immunoprecipitation Image 1: ZMYND8 (E2Z4U) Rabbit mAb Expand Image
使用 SimpleChIP Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005 对 ZR-75-1 细胞的交联染色质与 ZMYND8 (E2Z4U) Rabbit mAb 进行染色质免疫沉淀分析。使用 DNA Library Prep Kit for Illumina #56795 制备 DNA 库。本图显示了 ZMYND8 的已知靶基因 ACTB 内的结合(参见包含 ChIP-qPCR 数据的其他结果图)。
Chromatin Immunoprecipitation Image 2: ZMYND8 (E2Z4U) Rabbit mAb Expand Image
使用 SimpleChIP Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005 对 ZR-75-1 细胞的交联染色质与 ZMYND8 (E2Z4U) Rabbit mAb 进行染色质免疫沉淀分析。使用 DNA Library Prep Kit for Illumina #56795 制备 DNA 库。结果图显示在染色体7(上图)内的结合,包括 ZMYND8 的已知靶基因 ACTB (下图)(参见包含 ChIP-qPCR 数据的其他结果图)。
Chromatin Immunoprecipitation Image 3: ZMYND8 (E2Z4U) Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005 对 ZR-75-1 细胞的交联染色质与 ZMYND8 (E2Z4U) Rabbit mAb 或 Normal Rabbit IgG #2729 进行染色质免疫沉淀。使用 human S100A2 exon 1 primers、human FxyD3 intron 3 primers 和 SimpleChIP® Human α Satellite Repeat Primers #4486 进行实时 PCR 来对富集的 DNA 进行定量。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。