Revision 1

#55962Store 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

REACTIVITY:

H Mk

SENSITIVITY:

Endogenous

MW (kDa):

42

Source/Isotype:

Rabbit IgG

UniProt ID:

#P28356

Entrez-Gene Id:

3235

Product Information

Product Usage Information

For optimal ChIP 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

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

HOXD9 (E3F2T) Rabbit mAb recognizes endogenous levels of total HOXD9 protein. This antibody does not cross-react with HOXA9, HOXB9 or HOXC9 proteins. This antibody also weakly recognizes non-specific bands of unknown origin at 55 and 70 kDa in some cell lines.

Species Reactivity:

Human, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly159 of human HOXD9 protein.

Background

Homeobox protein Hox-D9 (HOXD9) is a sequence-specific transcription factor that is part of a developmental regulatory program that provides cells with specific positional identities on the anterior-posterior axis. HOXD9 is developmentally expressed in structures of either mesodermal or neuro-ectodermal origin, such as developing limbs, gonads, and the central nervous system (1-6). HOXD9 plays a critical role in regulation of limb development, neuronal development, and development of mammary glands and gonads in many organisms (1-6). The HOXD9 gene promoter is found to be hypermethylated and silenced in multiple types of cancer, including breast cancer, melanoma brain metastases, and cholangiocarcinomas (7-9). In addition, HOXD9 expression is increased in other types of cancer, including human glioblastomas and astrocytomas, where expression appears to drive growth of the tumors (10).

  1. Raines, A.M. et al. (2013) Development 140, 2942-52.
  2. Wang, Z. et al. (2010) BMC Genomics 11, 619.
  3. Freitas, R. et al. (2007) PLoS One 2, e754.
  4. Tanaka, M. et al. (2005) Dev Biol 281, 227-39.
  5. de la Cruz, C.C. et al. (1999) Dev Biol 216, 595-610.
  6. Chen, F. and Capecchi, M.R. (1999) Proc Natl Acad Sci U S A 96, 541-6.
  7. Makiyama, K. et al. (2005) Oncol Rep 13, 673-9.
  8. Marzese, D.M. et al. (2014) Hum Mol Genet 23, 226-38.
  9. Sriraksa, R. et al. (2013) Cancer Prev Res (Phila) 6, 1348-55.
  10. Tabuse, M. et al. (2011) Mol Cancer 10, 60.

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

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 1
#55962

HOXD9 (E3F2T) Rabbit mAb

Western Blotting Image 1: HOXD9 (E3F2T) Rabbit mAb Expand Image
使用 HOXD9 (E3F2T) Rabbit mAb 对不同细胞系的提取物进行蛋白印迹分析。
Western Blotting Image 2: HOXD9 (E3F2T) Rabbit mAb Expand Image
使用 HOXD9 (E3F2T) Rabbit mAb(上图)或 DYKDDDDK Tag (D6W5B) Rabbit mAb #14793(下图)对转染空载 (-) 或转染表达带有 Myc/DDK 标签的全长人 HOXA9 蛋白 (hHOXA9-Myc/DDK; +)、带有 Myc/DDK 标签的全长人 HOXC9 蛋白 (hHOXC9-Myc/DDK; +) 和带有 Myc/DDK 标签的全长人 HOXD9 蛋白 (hHOXD9-Myc/DDK; +) 的表达载体的 293T 细胞的提取物进行蛋白印迹分析。
Immunoprecipitation Image 1: HOXD9 (E3F2T) Rabbit mAb Expand Image
对 A-673 细胞提取物 HOXD9 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 HOXD9 (E3F2T) Rabbit mAb。使用 HOXD9 (E3F2T) Rabbit mAb 进行蛋白印迹分析。小鼠Anti-兔 IgG (轻链特异) (D4W3E) 单克隆抗体 (HRP 偶联) #93702 用作二抗。
Chromatin Immunoprecipitation Image 1: HOXD9 (E3F2T) Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005 对未经处理的 A-673 细胞的交联染色质与 EOMES (D8D1R) Rabbit mAb 或 Normal Rabbit IgG #2729 进行染色质免疫沉淀分析。使用 SimpleChIP® Human KLF3 Intron 1 Primers #23707, human EHD3 downstream primers 和 SimpleChIP® Human α Satellite Repeat Primers #4486 并通过实时 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。