Revision 4

#83527Store 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

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

70

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q9GZV8

Entrez-Gene Id:

63978

Product Information

Product Usage Information

For optimal ChIP and ChIP-seq results, use 10 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.

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Immunofluorescence (Immunocytochemistry) 1:400
Flow Cytometry (Fixed/Permeabilized) 1:800
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

PRDM14 (E1D5S) XP® Rabbit mAb recognizes endogenous levels of total PRDM14 protein.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with full-length recombinant protein specific to human PRDM14 protein.

Background

PR domain zinc finger protein 14 (PRDM14) is a likely protein lysine methyltransferase that is primarily expressed in primordial germ cells and pluripotent embryonic stem cells. It is essential for the establishment and maintenance of primordial germ cells and critical for the maintenance of pluripotency in embryonic stem cells (1-3). PRDM14 represses genes involved in the differentiation of stem cells into various cell lineages, likely via a combination of interactions with TET proteins, the polycomb repressive complex 2 (PRC2), and CBFA2T2 (3-8). In addition, overexpression of PRDM14 in combination with Jarid2 promotes induced pluripotent stem cell (iPSC) formation (9). PRDM14 protein levels are overexpressed in certain cancers, including breast, leukemia (T-ALL), and non-small cell lung cancer (NSCLC) (10-13), and PRDM14 overexpression may serve as a novel prognostic marker in NSCLC (14). Targeting PRDM14 overexpression with a siRNA-based therapy was shown to decrease liver metastasis in a murine pancreatic cancer model, suggesting potential as a therapeutic option for cancers where this protein is abnormally expressed (15).

  1. Yamaji, M. et al. (2008) Nat Genet 40, 1016-22.
  2. Chia, N.Y. et al. (2010) Nature 468, 316-20.
  3. Tsuneyoshi, N. et al. (2008) Biochem Biophys Res Commun 367, 899-905.
  4. Ma, Z. et al. (2011) Nat Struct Mol Biol 18, 120-7.
  5. Okashita, N. et al. (2014) Development 141, 269-80.
  6. Yamaji, M. et al. (2013) Cell Stem Cell 12, 368-82.
  7. Nady, N. et al. (2015) Elife 4, e10150.
  8. Tu, S. et al. (2016) Nature 534, 387-90.
  9. Iseki, H. et al. (2016) Stem Cells 34, 322-33.
  10. Moelans, C.B. et al. (2010) Mod Pathol 23, 1029-39.
  11. Nishikawa, N. et al. (2007) Cancer Res 67, 9649-57.
  12. Carofino, B.L. et al. (2013) Dis Model Mech 6, 1494-506.
  13. Liu, B. et al. (2010) Zhongguo Fei Ai Za Zhi 13, 867-72.
  14. Zhang, T. et al. (2013) Med Oncol 30, 605.
  15. Moriya, C. et al. (2017) Carcinogenesis 38, 638-648.

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

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.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

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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
#83527

PRDM14 (E1D5S) XP® Rabbit mAb

Western Blotting Image 1: PRDM14 (E1D5S) XP® Rabbit mAb Expand Image
使用 PRDM14 (E1D5S) XP® 兔单克隆抗体(上图)和 GAPDH (D16H11) XP® 兔单克隆抗体#5174(下图)对不同细胞系的提取物进行蛋白印迹分析。正如预计的一样,MCF7 和 293T 细胞的 PRDM14 表达呈阴性。
Immunoprecipitation Image 1: PRDM14 (E1D5S) XP® Rabbit mAb Expand Image
对 NCCIT 细胞提取物 PRDM14 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 PRDM14 (E1D5S) XP® 兔单克隆抗体。使用 PRDM14 (E1D5S) XP® 兔单克隆抗体进行蛋白印迹分析。
Immunofluorescence Image 1: PRDM14 (E1D5S) XP® Rabbit mAb Expand Image
使用 PRDM14 (E1D5S) XP® 兔单克隆抗体(绿色)对 NCCIT 细胞(左图,阳性)和 293T 细胞(右图,阴性)进行共聚焦免疫荧光分析。肌动蛋白微丝由 DyLight 554 Phalloidin #13054(红色)标记。样本用 ProLong® Gold Antifade Reagent with DAPI #8961(蓝色)封片。
Flow Cytometry Image 1: PRDM14 (E1D5S) XP® Rabbit mAb Expand Image
使用 PRDM14 (E1D5S) XP® 兔单克隆抗体(实线)或浓度匹配的兔 (DA1E) 单克隆抗体 IgG XP® 同型对照 #3900(虚线)对 NCCIT 细胞(绿色)和 293T 细胞(蓝色)进行流式细胞分析。抗兔 IgG (H+L), F(ab')2 Fragment (Alexa Fluor®488 偶联物) #4412 用作二抗。
Chromatin Immunoprecipitation Image 1: PRDM14 (E1D5S) XP® 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 PRDM14 (E1D5S) XP® 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 PRDM14 的已知靶基因 POU5F1/Oct-4 内结合(参见包含 ChIP-qPCR 数据的其他结果图)。欲知其他 ChIP-seq 情况,请下载产品说明书。
Chromatin Immunoprecipitation Image 2: PRDM14 (E1D5S) XP® 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 PRDM14 (E1D5S) XP® 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 6(上图)内的结合,包括 PRDM14 的已知靶基因 POU5F1/Oct-4(下图)(参见包含 ChIP-qPCR 数据的其他结果图)。
Chromatin Immunoprecipitation Image 3: PRDM14 (E1D5S) XP® 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 NCCIT 细胞的交联染色质与 PRDM14 (E1D5S) XP® 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 Oct-4 启动子引物 #4641、人源 ASXL1 上游引物和 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。