Revision 4

#93801Store 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 M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

135, 140

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q9UPP1

Entrez-Gene Id:

23133

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 1:200
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

PHF8 (E6K3Y) Rabbit mAb recognizes endogenous levels of total PHF8 protein.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein containing Phe579 of human PHF8 protein.

Background

PHD finger protein 8 (PHF8) is a histone lysine demethylase that functions as a transcriptional activator by specifically demethylating a number of repressive histone methylation marks: mono- and di-methyl-histone H3 Lys9 (H3K9me1 and H3K9me2), di-methyl-histone H3 Lys27 (H3K27me2) and mono-methyl-histone H4 Lys20 (H4K20me1). PHF8 contains an N-terminal zinc finger-like PHD domain that binds tri-methylated histone H3 Lys4 (H3K4Me3) and a C-terminal jumonji domain that is responsible for the demethylase activity (1). Deletion and point mutations (F279S) in the jumonji domain of PHF8 are associated with the onset of X-linked mental retardation (XLMR). In addition, PHF8 is highly expressed in prostate cancer, laryngeal squamous cell carcinoma, and human non-small-cell lung cancer (NSCLC). Its expression is predictive of poor survival (2-4). Overexpression of PHF8 increases cell proliferation and cell motility, while silencing of PHF8 reduces cell proliferation, migration, and invasion (4).

  1. Horton, J.R. et al. (2010) Nat Struct Mol Biol 17, 38-43.
  2. Zhu, G. et al. (2015) Epigenomics 7, 143-53.
  3. Shen, Y. et al. (2014) Biochem Biophys Res Commun 451, 119-25.
  4. Björkman, M. et al. (2012) Oncogene 31, 3444-56.

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

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
#93801

PHF8 (E6K3Y) Rabbit mAb

Western Blotting Image 1: PHF8 (E6K3Y) Rabbit mAb Expand Image
使用 PHF8 (E6K3Y) 兔单克隆抗体 对不同细胞系的提取物进行蛋白印迹分析。
Immunoprecipitation Image 1: PHF8 (E6K3Y) Rabbit mAb Expand Image
对 MOLT-4 细胞提取物 PHF8 进行免疫沉淀。泳道 1 为 10% 输入,泳道 2 为兔 (DA1E) 单克隆抗体 IgG XP® 同型对照 #3900,泳道 3 为 PHF8 (E6K3Y) 兔单克隆抗体。使用 PHF8 (E6K3Y) 兔单克隆抗体 进行蛋白印迹分析。
Chromatin Immunoprecipitation Image 1: PHF8 (E6K3Y) Rabbit mAb Expand Image
使用 SimpleChIP Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005 对 SH-SY5Y 细胞的交联染色质与 PHF8 (E6K3Y) 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 KDM5C 基因内结合。欲知其他 ChIP-seq 情况,请下载产品说明书。
Chromatin Immunoprecipitation Image 2: PHF8 (E6K3Y) Rabbit mAb Expand Image
使用 SimpleChIP Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005 对 SH-SY5Y 细胞的交联染色质与 PHF8 (E6K3Y) 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 X(上图)内的结合,包括 KDM5C 基因(下图)。
Chromatin Immunoprecipitation Image 3: PHF8 (E6K3Y) Rabbit mAb Expand Image
使用 SimpleChIP® 酶解染色质免疫共沉淀试剂盒(磁珠法) #9005 对 SH-SY5Y 细胞的交联染色质与 PHF8 (E6K3Y) 兔单克隆抗体或正常兔 IgG #2729 进行染色质免疫沉淀。使用 SimpleChIP® 人源 JARID1C 外显子 1 引物 #52773、人源 ZNF41 外显子 1 引物、人源 ZNF81 启动子引物 和 SimpleChIP® 人源 α 卫星重复序列引物 #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。