Revision 4

#65127Store 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, C&R

REACTIVITY:

H Mk

SENSITIVITY:

Endogenous

MW (kDa):

66, 152

Source/Isotype:

Rabbit IgG

UniProt ID:

#O96028

Entrez-Gene Id:

7468

Product Information

Product Usage Information

The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.
Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:200
CUT&RUN 1:25

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

WHSC1 (D4Z8Z) Rabbit mAb recognizes endogenous levels of total WHSC1 protein. Based on antigen sequence, this antibody should detect WHSC1 Isoform II (UniProt O96028-1) and REIIBP (UniProt O96028-4). This antibody does not detect Isoform I (UniProt O96028-3).

Species Reactivity:

Human, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the carboxy terminus of human WHSC1 protein.

Background

Wolf-Hirschhorn syndrome candidate protein 1 (WHSC1), also known as Nuclear SET domain-containing protein 2 (NSD2) and Multiple myeloma SET domain-containing protein (MMSET), is a histone methyltransferase protein that catalyzes the di-methylation of histone H3 Lys36, a mark that is associated with transcriptionally active regions of the genome (1,2). WHSC1 haploinsufficiency is implicated in the developmental disorder known as Wolf-Hirschhorn syndrome (WHS), which is characterized by growth and mental retardation, and congenital heart defects (3). In addition, WHSC1 is overexpressed and associated with poor prognosis in a large variety of human cancers, including neuroblastoma, and ovarian, hepatocellular, endometrial, and colorectal carcinoma (4-8). In multiple myeloma, 15-20% of all patients show overexpression of WHSC1 due to a t(4;14)(p16.3;q32) chromosomal translocation that places the WHSC1 gene upstream of a strong enhancer (2). The WHSC1 gene encodes for three major protein isoforms. Isoform II is the full-length protein containing 1365 amino acids. Isoform I, which is generated by alternative splicing, is identical to the N-terminal 647 amino acids of isoform II. A third isoform, known as REIIBP, is generated from a transcript initiating in intron 9 of the WHSC1 locus and is identical to the C-terminal 584 amino acids of isoform II.

  1. Vougiouklakis, T. et al. (2015) Epigenomics 7, 863-74.
  2. Dolloff, N.G. and Talamo, G. (2013) Adv Exp Med Biol 779, 197-221.
  3. Stec, I. et al. (1998) Hum Mol Genet 7, 1071-82.
  4. Hudlebusch, H.R. et al. (2011) Clin Cancer Res 17, 2919-33.
  5. Yang, S. et al. (2013) Biomarkers 18, 257-63.
  6. Zhou, P. et al. (2013) Pathol Oncol Res 19, 303-9.
  7. Xiao, M. et al. (2013) J Surg Oncol 107, 428-32.
  8. Hudlebusch, H.R. et al. (2011) Cancer Res 71, 4226-35.

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 C&R: CUT&RUN

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
#65127

WHSC1 (D4Z8Q) Rabbit mAb

Western Blotting Image 1: WHSC1 (D4Z8Q) Rabbit mAb Expand Image
使用 WHSC1 (D4Z8Q) 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: WHSC1 (D4Z8Q) Rabbit mAb Expand Image
对 MOLT-4 细胞提取物 WHSC1 进行免疫沉淀。泳道 1 为 10% 敲入,泳道 2 为兔 (DA1E) 单克隆抗体 IgG XP® 同型对照 #3900,泳道 3 为 WHSC1 (D4Z8Q) 兔单克隆抗体。使用 WHSC1 (D4Z8Q) 兔单克隆抗体 进行蛋白印迹分析。
CUT and RUN Image 1: WHSC1 (D4Z8Q) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652 对经 TNFa(30 ng/ml,1 小时)和 WHSC1 (D4Z8Q) Rabbit mAb 处理的 HeLa 细胞进行 CUT&RUN 检测。使用 DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795 制备 DNA 文库。该图显示在 ARID1A 基因内结合。
CUT and RUN Image 2: WHSC1 (D4Z8Q) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652 对经 TNFa(30 ng/ml,1 小时)和 WHSC1 (D4Z8Q) Rabbit mAb 处理的 HeLa 细胞进行 CUT&RUN 检测。使用 DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。该图显示在染色体 1(上图)内的结合,包括 ARID1A 基因(下图)。
CUT and RUN Image 3: WHSC1 (D4Z8Q) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652 对经人 TNF-α #8902(30 ng/ml,1 小时)和 WHSC1 (D4Z8Q) Rabbit mAb 或 Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362 处理的 HeLa 细胞进行 CUT&RUN 检测。使用 SimpleChIP® Human IκBα Promoter Primers #5552、human IL-8 promoter primers 和 SimpleChIP® Human α Satellite Repeat Primers #4486,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。